NYMU Taipei/Benchman

From 2007.igem.org

(Difference between revisions)
Line 1: Line 1:
-
 
+
<h3>OD and GEL checks</h3>
 +
<table width="100%" align="center" border="1">
 +
    <tbody>
 +
        <tr align="center">
 +
            <td><font size="1"><strong>Part name<br />
 +
            </strong></font></td>
 +
            <td><font size="1"><strong>Type and Description<br />
 +
            </strong></font></td>
 +
            <td><font size="1"><strong>length (bp) </strong></font></td>
 +
            <td><font size="1"><strong>PCR&nbsp; check</strong></font></td>
 +
            <td><font size="1"><strong>weight (g) </strong></font></td>
 +
            <td><font size="1"><strong>concentration (ug/uL) </strong></font></td>
 +
            <td><font size="1"><strong>A260/A280</strong></font></td>
 +
            <td><font size="1"><strong>A260/A230</strong></font></td>
 +
            <td><font size="1"><strong>date</strong></font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td align="left"><font size="1">pOmpC (P.E. #1)</font></td>
 +
            <td align="left"><font size="1">plasmid extraction #1</font></td>
 +
            <td align="right"><font size="1">2,271</font></td>
 +
            <td align="right">&nbsp;</td>
 +
            <td align="right"><font size="1">&nbsp;</font></td>
 +
            <td align="right"><font size="1">0.20</font></td>
 +
            <td align="right"><font size="1">1.30</font></td>
 +
            <td align="right"><font size="1">0.97</font></td>
 +
            <td align="right"><font size="1"><a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_3">Oct 03, 07</a></font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td align="left"><font size="1">pOmpC (P.E. #2)</font></td>
 +
            <td align="left"><font size="1">plasmid extraction #2</font></td>
 +
            <td align="right"><font size="1">2,271</font></td>
 +
            <td align="right">&nbsp;</td>
 +
            <td align="right"><font size="1">&nbsp;</font></td>
 +
            <td align="right"><font size="1">0.19</font></td>
 +
            <td align="right"><font size="1">1.33</font></td>
 +
            <td align="right"><font size="1">1.00</font></td>
 +
            <td align="right"><font size="1"><a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_3">Oct 03, 07</a></font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td align="left"><font size="1">pOmpC (P.E. #3)</font></td>
 +
            <td align="left"><font size="1">plasmid extraction #3</font></td>
 +
            <td align="right"><font size="1">2,271</font></td>
 +
            <td align="right">&nbsp;</td>
 +
            <td align="right"><font size="1">&nbsp;</font></td>
 +
            <td align="right"><font size="1">0.21</font></td>
 +
            <td align="right"><font size="1">1.42</font></td>
 +
            <td align="right"><font size="1">1.12</font></td>
 +
            <td align="right"><font size="1"><a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_3">Oct 03, 07</a></font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td align="left"><font size="1">pOmpC (P.E. #4)</font></td>
 +
            <td align="left"><font size="1">plasmid extraction #4</font></td>
 +
            <td align="right"><font size="1">2,271</font></td>
 +
            <td align="right">&nbsp;</td>
 +
            <td align="right"><font size="1">&nbsp;</font></td>
 +
            <td align="right"><font size="1">0.25</font></td>
 +
            <td align="right"><font size="1">1.37</font></td>
 +
            <td align="right"><font size="1">1.03</font></td>
 +
            <td align="right"><font size="1"><a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_3">Oct 03, 07</a></font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td align="left"><font size="1">pOmpC (D. SP)</font></td>
 +
            <td align="left"><font size="1">digestion (S. P.) as back vector, #4 selected</font></td>
 +
            <td align="right"><font size="1">2,253</font></td>
 +
            <td align="right">&nbsp;</td>
 +
            <td align="right"><font size="1">0.14</font></td>
 +
            <td align="right"><font size="1">0.01</font></td>
 +
            <td align="right"><font size="1">n/a</font></td>
 +
            <td align="right"><font size="1">n/a</font></td>
 +
            <td align="right"><font size="1"><a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_4">Oct 04, 07</a></font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td align="left"><font size="1">TATA_INSA (D. XP)</font></td>
 +
            <td align="left"><font size="1">digestion (X. P.) as back insert</font></td>
 +
            <td align="right"><font size="1">&nbsp;193</font></td>
 +
            <td align="right">&nbsp;</td>
 +
            <td align="right"><font size="1">0.25</font></td>
 +
            <td align="right"><font size="1">0.02</font></td>
 +
            <td align="right"><font size="1">n/a</font></td>
 +
            <td align="right"><font size="1">0.29</font></td>
 +
            <td align="right"><font size="1"><a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_3">Oct 03, 07</a></font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td align="left"><font size="1">TATA_INSA (PCR)</font></td>
 +
            <td align="left"><font size="1">PCR for TATA + INSA</font></td>
 +
            <td align="right"><font size="1">209</font></td>
 +
            <td align="right">&nbsp;</td>
 +
            <td align="right"><font size="1">0.24; 0.30</font></td>
 +
            <td align="right"><font size="1">&nbsp;</font></td>
 +
            <td align="right"><font size="1">&nbsp;</font></td>
 +
            <td align="right"><font size="1">&nbsp;</font></td>
 +
            <td align="right"><font size="1">Oct 9, 07</font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td align="left"><font size="1">TATB_INSB (PCR)</font></td>
 +
            <td align="left"><font size="1">PCR for TATB + INSB</font></td>
 +
            <td align="right"><font size="1">233</font></td>
 +
            <td align="right">&nbsp;</td>
 +
            <td align="right"><font size="1">0.35; 0.35</font></td>
 +
            <td align="right"><font size="1">&nbsp;</font></td>
 +
            <td align="right"><font size="1">&nbsp;</font></td>
 +
            <td align="right"><font size="1">&nbsp;</font></td>
 +
            <td align="right"><font size="1">Oct 9, 07</font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td style="background-color: rgb(255,255,153)" align="left"><font size="1">TATA_INSA (D. XP)</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="left"><font size="1">digestion (X. P.) as <strong>back insert</strong></font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">&nbsp;193</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right">&nbsp;</td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">n/a</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">n/a</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">n/a</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1"><a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_10">Oct 10, 07</a></font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td style="background-color: rgb(255,255,153)" align="left"><font size="1">TATB_INSB (D. XP)</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="left"><font size="1">digestion (X. P.) as <strong>back insert</strong></font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">220</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right">&nbsp;</td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">n/a</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">n/a</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">n/a</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1"><a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_10">Oct 10, 07</a></font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td style="background-color: rgb(255,255,153)" align="left"><font size="1">pOmpC (D. SP)</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="left"><font size="1">digestion (S. P.) as <strong>back vector</strong>, #1 selected</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">2,253</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right">&nbsp;</td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">n/a</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">n/a</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">n/a</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">Oct 11, 07</font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td style="background-color: rgb(204,255,255)" align="left"><font size="1">pOmpC + TATA_INSA</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="left"><font size="1">back ligation pOmpC (D. SP) + TATA_INSA (D. XP)</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;2,454</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1"><font color="#ff0000">430</font>, 613</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right">&nbsp;</td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">Oct 13, 07 &amp; Oct 16,07</font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td style="background-color: rgb(204,255,255)" align="left"><font size="1">pOmpC+TATA_INSA (D. SP)</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="left"><font size="1">digestion (S. P.) as <strong>back vector</strong></font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">2,436 2,427</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right">&nbsp;</td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td style="background-color: rgb(204,255,255)" align="left"><font size="1">pOmpC+TATA_INSA + TATB_INSB</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="left"><font size="1">back ligation pOmpC+TATA_INSA (D. SP) + TATB_INSB (D. XP)</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">2,655</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font color="#ff0000" size="1">613</font><font size="1">, 814</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td style="background-color: rgb(255,204,153)" align="left"><font size="1">TATB_INSB (D. ES)</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="left"><font size="1">digestion (E. S.) as front insert (</font><font color="#ff0000" size="1">wrong!!!</font><font size="1">)</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right">&nbsp;</td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">0.27</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">0.03</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">n/a</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">0.21</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1"><a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_3">Oct 03, 07</a></font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td style="background-color: rgb(255,204,153)" align="left"><font size="1">CinR+HSL+D-term (P.E.)</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="left"><font size="1">plasmid extraction #1, #3, #4 (</font><font color="#ff0000" size="1">wrong!!!</font><font size="1">)</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">&nbsp;4,804</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right">&nbsp;</td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">0.41, 0.25, 0.41</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1"><a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_9_27#CinR.2BHSL.2BD-term_check">Sep 27, 07</a></font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td style="background-color: rgb(255,204,153)" align="left"><font size="1">CinR+HSL+D-term (D. E)</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="left"><font size="1">first digestion (E.) as front vector (</font><font color="#ff0000" size="1">wrong!!!</font><font size="1">)</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">4,804</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right">&nbsp;</td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">0.19</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1"><a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_3">Oct 03, 07</a></font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td style="background-color: rgb(255,204,153)" align="left"><font size="1">CinR+HSL+D-term (D. EX)</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="left"><font size="1">second digestion (X.) as front vector (</font><font color="#ff0000" size="1">wrong!!!</font><font size="1">)</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">4,804</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right">&nbsp;</td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">0.19</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">0.02</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">n/a</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">0.13</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1"><a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_4">Oct 04, 07</a></font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td style="background-color: rgb(255,204,153)" align="left"><font size="1">TATB_INSB + CinR+HSL+D-term</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="left"><font size="1">front ligation TATB_INSB (D. ES) + CinR+HSL+D-term (D. EX) (</font><font color="#ff0000" size="1">wrong!!!</font><font size="1">)</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right">&nbsp;</td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">Oct 05, 07</font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td style="background-color: rgb(204,255,255)" align="center"><font size="1">pOmpC+TATA_INSA+TATB_INSB (D. SP)</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="left"><font size="1">digestion (S.P.) as <strong>back vector</strong></font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">2,637</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right">&nbsp;</td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td style="background-color: rgb(204,255,255)" align="center"><font size="1">CinR+HSL+D-term (D. XP)</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="left"><font size="1">digestion (X.P.) as <strong>back insert</strong></font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">1,647</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right">&nbsp;</td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td align="center"><font size="1"><strong>system 1</strong></font></td>
 +
            <td align="left"><font size="1">ligation pOmpC+TATA_INSA+TATB_INSB + CinR+HSL+D-term</font></td>
 +
            <td align="right"><font size="1">4,292</font></td>
 +
            <td align="right"><font color="#ff0000" size="1">814</font><font size="1">, 2,451</font></td>
 +
            <td align="right"><font size="1">&nbsp;</font></td>
 +
            <td align="right"><font size="1">&nbsp;</font></td>
 +
            <td align="right"><font size="1">&nbsp;</font></td>
 +
            <td align="right"><font size="1">&nbsp;</font></td>
 +
            <td align="right"><font size="1">&nbsp;</font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td align="left"><font size="1">pCinRHSL (P.E. #1)</font></td>
 +
            <td align="left"><font size="1">plasmid extraction #1</font></td>
 +
            <td align="right"><font size="1">&nbsp;</font></td>
 +
            <td align="right">&nbsp;</td>
 +
            <td align="right"><font size="1">&nbsp;</font></td>
 +
            <td align="right"><font size="1">0.23</font></td>
 +
            <td align="right"><font size="1">1.70</font></td>
 +
            <td align="right"><font size="1">1.49</font></td>
 +
            <td align="right"><font size="1"><a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_3">Oct 03, 07</a></font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td align="left"><font size="1">pCinRHSL (P.E. #2)</font></td>
 +
            <td align="left"><font size="1">plasmid extraction #2</font></td>
 +
            <td align="right"><font size="1">&nbsp;</font></td>
 +
            <td align="right">&nbsp;</td>
 +
            <td align="right"><font size="1">&nbsp;</font></td>
 +
            <td align="right"><font size="1">0.29</font></td>
 +
            <td align="right"><font size="1">1.58</font></td>
 +
            <td align="right"><font size="1">1.26</font></td>
 +
            <td align="right"><font size="1"><a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_3">Oct 03, 07</a></font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td align="left"><font size="1">pCinRHSL (D. SP)</font></td>
 +
            <td align="left"><font size="1">digestion (S. P.) as back vector, #1 selected</font></td>
 +
            <td align="right"><font size="1">&nbsp;</font></td>
 +
            <td align="right">&nbsp;</td>
 +
            <td align="right"><font size="1">0.17</font></td>
 +
            <td align="right"><font size="1">0.03</font></td>
 +
            <td align="right"><font size="1">n/a</font></td>
 +
            <td align="right"><font size="1">0.18</font></td>
 +
            <td align="right"><font size="1"><a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_4">Oct 04, 07</a></font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td align="left"><font size="1">OmpRBS (D. XP)</font></td>
 +
            <td align="left"><font size="1">digestion (X. P.) as back insert</font></td>
 +
            <td align="right"><font size="1">&nbsp;</font></td>
 +
            <td align="right">&nbsp;</td>
 +
            <td align="right"><font size="1">0.24</font></td>
 +
            <td align="right"><font size="1">0.01</font></td>
 +
            <td align="right"><font size="1">n/a</font></td>
 +
            <td align="right"><font size="1">n/a</font></td>
 +
            <td align="right"><font size="1"><a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_3">Oct 03, 07</a></font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td style="background-color: rgb(255,255,153)" align="left"><font size="1">pCinRHSL+OmpRBS</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="left"><font size="1">ligation pCinRHSL (D. SP) + OmpRBS (D. XP)</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right">&nbsp;</td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">Oct 05, 07</font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td style="background-color: rgb(255,204,153)" align="left"><font size="1">pCinRHSL+OmpRBS (D. SP)</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="left"><font size="1">digestion (S. P.) pCinRHSL+OmpRBS as back vector (a bit smear)</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">2,443</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right">&nbsp;</td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">0.14 (too few)</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">Oct 9, 07</font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td style="background-color: rgb(255,255,153)" align="left"><font size="1">pCinRHSL+OmpRBS (plasmid)</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="left"><font size="1">plasmid extraction of pCinRHSL+OmpRBS (clone 7 and clone 10 on the plate) after liquid culture</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right">&nbsp;</td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">0.020; 0.019</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">1.95; 1.83</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">2.18; 2.31</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">Oct 10, 07</font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td style="background-color: rgb(255,204,153)" align="left"><font size="1">pCinRHSL+OmpRBS (D. SP)</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="left"><font size="1">digestion (S. P.) pCinRHSL+OmpRBS as back vector (smear)</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">&nbsp;2,443</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right">&nbsp;</td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">Oct 10, 07</font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td style="background-color: rgb(255,255,153)" align="left"><font size="1">pCinRHSL+OmpRBS (D. SP)</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="left"><font size="1">digestion (S. P.) pCinRHSL+OmpRBS as <strong>back vector</strong> using plasmid from clone 7</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">2,443</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right">&nbsp;</td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">0.05</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">n/a</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">0.22</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">Oct 11, 07</font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td style="background-color: rgb(255,204,153)" align="left"><font size="1">TATD_IDE (D. XP)</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="left"><font size="1">digestion (X. P.) as back insert</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">3,190</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right">&nbsp;</td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">0.10 (too few)</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,204,153)" align="right"><font size="1">Oct 9, 07</font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td style="background-color: rgb(255,255,153)" align="left"><font size="1">TATD + IDE (PCR)</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="left"><font size="1">re-PCR for TATD + IDE</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">3,203</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right">&nbsp;</td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(255,255,153)" align="right"><font size="1">Oct 10, 07</font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td style="background-color: rgb(204,255,255)" align="left"><font size="1">TATD_IDE (D. XP)</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="left"><font size="1">digestion (X. P.) as <strong>back insert</strong></font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;3,190</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right">&nbsp;</td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">Oct 11, 07</font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td style="background-color: rgb(204,255,255)" align="left"><font size="1">pCinRHSL+OmpRBS + TATD_IDE</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="left"><font size="1">ligation pCinRHSL+OmpRBS + TATD_IDE</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right">&nbsp;</td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td style="background-color: rgb(204,255,255)" align="left"><font size="1">pCinRHSL+OmpRBS+TATD_IDE (D. SP)</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="left"><font size="1">digestion (S. P.) as <strong>back vector</strong></font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right">&nbsp;</td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td style="background-color: rgb(204,255,255)" align="left"><font size="1">D-term (D. XP)</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="left"><font size="1">digestion (X. P.) as <strong>back insert</strong></font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right">&nbsp;</td>
 +
            <td style="background-color: rgb(204,255,255)"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
            <td style="background-color: rgb(204,255,255)" align="right"><font size="1">&nbsp;</font></td>
 +
        </tr>
 +
        <tr align="center">
 +
            <td align="center"><font size="1"><strong>system 2</strong></font></td>
 +
            <td align="left"><font size="1">ligation pCinRHSL+OmpRBS+TATD_IDE + D-term</font></td>
 +
            <td align="right"><font size="1">&nbsp;</font></td>
 +
            <td align="right">&nbsp;</td>
 +
            <td><font size="1">&nbsp;</font></td>
 +
            <td><font size="1">&nbsp;</font></td>
 +
            <td align="right"><font size="1">&nbsp;</font></td>
 +
            <td align="right"><font size="1">&nbsp;</font></td>
 +
            <td align="right"><font size="1">&nbsp;</font></td>
 +
        </tr>
 +
    </tbody>
 +
</table>
 +
<hr />
 +
<h3>Notes</h3>
 +
<ul>
 +
    <li>A260/A280 should be around 1.70</li>
 +
    <li>A260/A230 should be less than 3.00</li>
 +
    <li>color key
 +
    <ul>
 +
        <li>problemistic (read)</li>
 +
        <li>finished (yellow)</li>
 +
        <li>await (blue)</li>
 +
    </ul>
 +
    </li>
 +
    <li>digestion and buffer
 +
    <ul>
 +
        <li>SepI and PstI (<strong>SP</strong>) digestion using buffer <strong>2</strong>, creates back <strong>vector</strong></li>
 +
        <li>XbaI and PstI (<strong>XP</strong>) digestion using buffer <strong>3</strong>, creates back <strong>insert</strong></li>
 +
    </ul>
 +
    </li>
 +
    <li>Assembly process
 +
    <ul>
 +
        <li><strong>STEP 1</strong>. prepare <strong>insert and vector</strong> by <strong>digestion</strong></li>
 +
        <li><strong>STEP 2</strong>. check digestion by GEL separation
 +
        <ul>
 +
            <li>if band is clear at correct position (length), cut the gel and purify/extract it</li>
 +
            <li>else go back to STEP 1.</li>
 +
        </ul>
 +
        </li>
 +
        <li><strong>STEP 3</strong>. <strong>ligate </strong>insert with vector, <strong>transform </strong>it into competent cell, and extract plasmid from liquid culture</li>
 +
        <li><strong>STEP 4</strong>. check ligation by <strong>VF2, VR PCR</strong> with GEL separation
 +
        <ul>
 +
            <li>if band is clear at correct position (length), cut the gel and purify/extract it (ligation is done)</li>
 +
            <li>else go back to STEP 3.</li>
 +
        </ul>
 +
        </li>
 +
    </ul>
 +
    </li>
 +
</ul>
[https://2007.igem.org/NYMU_Taipei/Welcome Back]
[https://2007.igem.org/NYMU_Taipei/Welcome Back]

Revision as of 13:37, 18 October 2007

OD and GEL checks

<tbody> </tbody>
Part name
 Type and Description
 length (bp) PCR  check weight (g) concentration (ug/uL) A260/A280 A260/A230 date
pOmpC (P.E. #1) plasmid extraction #1 2,271     0.20 1.30 0.97 <a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_3">Oct 03, 07</a>
pOmpC (P.E. #2) plasmid extraction #2 2,271     0.19 1.33 1.00 <a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_3">Oct 03, 07</a>
pOmpC (P.E. #3) plasmid extraction #3 2,271     0.21 1.42 1.12 <a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_3">Oct 03, 07</a>
pOmpC (P.E. #4) plasmid extraction #4 2,271     0.25 1.37 1.03 <a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_3">Oct 03, 07</a>
pOmpC (D. SP) digestion (S. P.) as back vector, #4 selected 2,253   0.14 0.01 n/a n/a <a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_4">Oct 04, 07</a>
TATA_INSA (D. XP) digestion (X. P.) as back insert  193   0.25 0.02 n/a 0.29 <a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_3">Oct 03, 07</a>
TATA_INSA (PCR) PCR for TATA + INSA 209   0.24; 0.30       Oct 9, 07
TATB_INSB (PCR) PCR for TATB + INSB 233   0.35; 0.35       Oct 9, 07
TATA_INSA (D. XP) digestion (X. P.) as back insert  193     n/a n/a n/a <a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_10">Oct 10, 07</a>
TATB_INSB (D. XP) digestion (X. P.) as back insert 220     n/a n/a n/a <a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_10">Oct 10, 07</a>
pOmpC (D. SP) digestion (S. P.) as back vector, #1 selected 2,253     n/a n/a n/a Oct 11, 07
pOmpC + TATA_INSA back ligation pOmpC (D. SP) + TATA_INSA (D. XP)  2,454 430, 613         Oct 13, 07 & Oct 16,07
pOmpC+TATA_INSA (D. SP) digestion (S. P.) as back vector 2,436 2,427            
pOmpC+TATA_INSA + TATB_INSB back ligation pOmpC+TATA_INSA (D. SP) + TATB_INSB (D. XP) 2,655 613, 814          
TATB_INSB (D. ES) digestion (E. S.) as front insert (wrong!!!)     0.27 0.03 n/a 0.21 <a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_3">Oct 03, 07</a>
CinR+HSL+D-term (P.E.) plasmid extraction #1, #3, #4 (wrong!!!)  4,804     0.41, 0.25, 0.41     <a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_9_27#CinR.2BHSL.2BD-term_check">Sep 27, 07</a>
CinR+HSL+D-term (D. E) first digestion (E.) as front vector (wrong!!!) 4,804   0.19       <a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_3">Oct 03, 07</a>
CinR+HSL+D-term (D. EX) second digestion (X.) as front vector (wrong!!!) 4,804   0.19 0.02 n/a 0.13 <a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_4">Oct 04, 07</a>
TATB_INSB + CinR+HSL+D-term front ligation TATB_INSB (D. ES) + CinR+HSL+D-term (D. EX) (wrong!!!)             Oct 05, 07
pOmpC+TATA_INSA+TATB_INSB (D. SP) digestion (S.P.) as back vector 2,637            
CinR+HSL+D-term (D. XP) digestion (X.P.) as back insert 1,647            
system 1 ligation pOmpC+TATA_INSA+TATB_INSB + CinR+HSL+D-term 4,292 814, 2,451          
pCinRHSL (P.E. #1) plasmid extraction #1       0.23 1.70 1.49 <a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_3">Oct 03, 07</a>
pCinRHSL (P.E. #2) plasmid extraction #2       0.29 1.58 1.26 <a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_3">Oct 03, 07</a>
pCinRHSL (D. SP) digestion (S. P.) as back vector, #1 selected     0.17 0.03 n/a 0.18 <a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_4">Oct 04, 07</a>
OmpRBS (D. XP) digestion (X. P.) as back insert     0.24 0.01 n/a n/a <a href="https://2007.igem.org/NYMU_Taipei/Lab_Notes/2007_10_3">Oct 03, 07</a>
pCinRHSL+OmpRBS ligation pCinRHSL (D. SP) + OmpRBS (D. XP)             Oct 05, 07
pCinRHSL+OmpRBS (D. SP) digestion (S. P.) pCinRHSL+OmpRBS as back vector (a bit smear) 2,443   0.14 (too few)       Oct 9, 07
pCinRHSL+OmpRBS (plasmid) plasmid extraction of pCinRHSL+OmpRBS (clone 7 and clone 10 on the plate) after liquid culture       0.020; 0.019 1.95; 1.83 2.18; 2.31 Oct 10, 07
pCinRHSL+OmpRBS (D. SP) digestion (S. P.) pCinRHSL+OmpRBS as back vector (smear)  2,443           Oct 10, 07
pCinRHSL+OmpRBS (D. SP) digestion (S. P.) pCinRHSL+OmpRBS as back vector using plasmid from clone 7 2,443     0.05 n/a 0.22 Oct 11, 07
TATD_IDE (D. XP) digestion (X. P.) as back insert 3,190   0.10 (too few)       Oct 9, 07
TATD + IDE (PCR) re-PCR for TATD + IDE 3,203           Oct 10, 07
TATD_IDE (D. XP) digestion (X. P.) as back insert  3,190           Oct 11, 07
pCinRHSL+OmpRBS + TATD_IDE ligation pCinRHSL+OmpRBS + TATD_IDE              
pCinRHSL+OmpRBS+TATD_IDE (D. SP) digestion (S. P.) as back vector              
D-term (D. XP) digestion (X. P.) as back insert              
system 2 ligation pCinRHSL+OmpRBS+TATD_IDE + D-term              

Notes

  • A260/A280 should be around 1.70
  • A260/A230 should be less than 3.00
  • color key
    • problemistic (read)
    • finished (yellow)
    • await (blue)
  • digestion and buffer
    • SepI and PstI (SP) digestion using buffer 2, creates back vector
    • XbaI and PstI (XP) digestion using buffer 3, creates back insert
  • Assembly process
    • STEP 1. prepare insert and vector by digestion
    • STEP 2. check digestion by GEL separation
      • if band is clear at correct position (length), cut the gel and purify/extract it
      • else go back to STEP 1.
    • STEP 3. ligate insert with vector, transform it into competent cell, and extract plasmid from liquid culture
    • STEP 4. check ligation by VF2, VR PCR with GEL separation
      • if band is clear at correct position (length), cut the gel and purify/extract it (ligation is done)
      • else go back to STEP 3.

Back

Retrieved from "http://2007.igem.org/wiki/index.php/NYMU_Taipei/Benchman"