Melbourne/Lab Notebook Weeks 13-16

From 2007.igem.org

(Difference between revisions)
(Week 13)
(Week 13)
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*# [[Melbourne/primary DNA marker|DNA ladder]]
*# [[Melbourne/primary DNA marker|DNA ladder]]
-
*# [[Melbourne/BBa_J61035|P4 8J]]-[[Melbourne/BBa_B0034|P1 3O]]-ComP 1 E/S
+
*# Gen-RBS-ComP 1 E/S
-
*# [[Melbourne/BBa_J61035|P4 8J]]-[[Melbourne/BBa_B0034|P1 3O]]-ComP 2 E/S
+
*# Gen-RBS-ComP 2 E/S
-
*# [[Melbourne/BBa_J61035|P4 8J]]-[[Melbourne/BBa_B0034|P1 3O]]-ComP 3 E/S
+
*# Gen-RBS-ComP 3 E/S
-
*# [[Melbourne/BBa_J61035|P4 8J]]-[[Melbourne/BBa_B0034|P1 3O]]-ComP 4 E/S
+
*# Gen-RBS-ComP 4 E/S
-
*# [[Melbourne/BBa_J61035|P4 8J]]-[[Melbourne/BBa_I15008|P2 21A]] 1 X/P
+
*# Gen-RBS-[[Melbourne/BBa_I15008|P2 21A]] 1 X/P
-
*# [[Melbourne/BBa_J61035|P4 8J]]-[[Melbourne/BBa_I15008|P2 21A]] 2 X/P
+
*# Gen-RBS-[[Melbourne/BBa_I15008|P2 21A]] 2 X/P
-
*# [[Melbourne/BBa_J61035|P4 8J]]-[[Melbourne/BBa_I15008|P2 21A]] 3 X/P
+
*# Gen-RBS-[[Melbourne/BBa_I15008|P2 21A]] 3 X/P
-
*# [[Melbourne/BBa_J61035|P4 8J]]-[[Melbourne/BBa_I15008|P2 21A]] 4 X/P
+
*# Gen-RBS-[[Melbourne/BBa_I15008|P2 21A]] 4 X/P
-
*# [[Melbourne/BBa_J61035|P4 8J]]-[[Melbourne/BBa_I15009|P2 21C]] 1 X/S
+
*# Gen-RBS-[[Melbourne/BBa_I15009|P2 21C]] 1 X/S
-
*# [[Melbourne/BBa_J61035|P4 8J]]-[[Melbourne/BBa_I15009|P2 21C]] 2 X/S
+
*# Gen-RBS-[[Melbourne/BBa_I15009|P2 21C]] 2 X/S
-
*# [[Melbourne/BBa_J61035|P4 8J]]-[[Melbourne/BBa_I15009|P2 21C]] 3 X/S
+
*# Gen-RBS-[[Melbourne/BBa_I15009|P2 21C]] 3 X/S
-
*# [[Melbourne/BBa_J61035|P4 8J]]-[[Melbourne/BBa_I15009|P2 21C]] 4 X/S
+
*# Gen-RBS-[[Melbourne/BBa_I15009|P2 21C]] 4 X/S
-
*# [[Melbourne/BBa_J61035|P4 8J]]-ComA 1 E/S
+
*# Gen-RBS-ComA 1 E/S
-
*# [[Melbourne/BBa_J61035|P4 8J]]-ComA 2 E/S
+
*# Gen-RBS-ComA 2 E/S
*# [[Melbourne/primary DNA marker|DNA ladder]]
*# [[Melbourne/primary DNA marker|DNA ladder]]
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*# [[Melbourne/primary DNA marker|DNA ladder]]
*# [[Melbourne/primary DNA marker|DNA ladder]]
*#
*#
-
*# [[Melbourne/BBa_J61035|P4 8J]]-ComP E/S 3.9kb
+
*# Gen-RBS-ComP E/S 3.9kb
*#
*#
-
*# [[Melbourne/BBa_J61035|P4 8J]]-ComA E/S 2.1kb  <font color=green> Had no DNA </font>
+
*# Gen-RBS-ComA E/S 2.1kb  <font color=green> Had no DNA </font>
*#
*#
-
*# [[Melbourne/BBa_J61035|P4 8J]]-[[Melbourne/BBa_I15008|P2 21A]] X/P 2.2kb  <font color=green> Took wrong fragment </font>
+
*# Gen-RBS-[[Melbourne/BBa_I15008|P2 21A]] X/P 2.2kb  <font color=green> Took wrong fragment </font>
*#
*#
-
*# [[Melbourne/BBa_J61035|P4 8J]]-[[Melbourne/BBa_I15009|P2 21C]] E/S 2.2kb
+
*# Gen-RBS-[[Melbourne/BBa_I15009|P2 21C]] E/S 2.2kb
*#
*#
*# L3 X/P 550bp  <font color=green> Ran off </font>
*# L3 X/P 550bp  <font color=green> Ran off </font>
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*# [[Melbourne/BBa_C0051|P1 5G]] E/X 3.3kb
*# [[Melbourne/BBa_C0051|P1 5G]] E/X 3.3kb
*# [[Melbourne/BBa_C0051|P1 5G]] E/X 3.3kb
*# [[Melbourne/BBa_C0051|P1 5G]] E/X 3.3kb
-
*#
 
*#
*#
*# [[Melbourne/primary DNA marker|DNA ladder]]
*# [[Melbourne/primary DNA marker|DNA ladder]]
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*[[Melbourne/Ligation Protocol|Ligated]] with 5min method:  
*[[Melbourne/Ligation Protocol|Ligated]] with 5min method:  
 +
*# Gen-RBS-ComP E/S into [[Melbourne/BBa_C0051|P1 5G]] E/X -survives on G/A
 +
*# Gen-RBS-[[Melbourne/BBa_I15009|P2 21C]] E/S into [[Melbourne/BBa_C0051|P1 5G]] E/X -survives on G/K
<font size=3><b>20 Sept 2007  
<font size=3><b>20 Sept 2007  
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</b>
</b>
</font><BR>
</font><BR>
 +
 +
*[[Melbourne/Transformation Protocol|Transformed]] into XL-10 gold cells using Quickchange Protocol.
==Week 14==
==Week 14==

Revision as of 13:10, 9 October 2007

<Return to Lab notebook> <team home page>

Contents

Week 13

16 Sept 2007


17 Sept 2007

Gel1:

    1. DNA ladder
    2. P17OA E/H
    3. P17OB E/H
    4. P1 5G A E/H
    5. P1 5G B E/H
    6. Omp-C1-reporter A X/P
    7. Omp-C1-reporter B X/P
    8. Omp-C1-reporter C X/P
    9. Omp-C1-reporter D X/P
    10. P2 21A A X/P
    11. P2 21A B X/P
    12. P2 21C A X/P
    13. P2 21C B X/P
    14. Death-ComA 5min ligation A X/P
    15. Death-ComA 5min ligation B X/P
    16. Death-ComA overnight ligation A X/P
    17. Death-ComA overnight ligation B X/P
    18. DNA ladder

Gel2:

    1. DNA ladder
    2. Death-ComP A X/P
    3. Death-ComP B X/P
    4. Cph8 A X/P
    5. Cph8 B X/P


-All had desired insert except for lanes 8-11 (OmpR-C1-reporter) in Gel 1 and lanes 4, 5 (CpH8) in Gel 2.


18 Sept 2007

  • Liquid cultured 4 colonies each from transformants of 17/9 (2 from Death-ComA).


19 Sept 2007

-Ran at 100V for 45min. -Lanes 4, 5, 7, 8, 9, 10, 11, 12, 13, 14, and 15 had desired bands.


    1. Gen-RBS-ComP E/S into P1 5G E/X -survives on G/A
    2. Gen-RBS-P2 21C E/S into P1 5G E/X -survives on G/K

20 Sept 2007


21 Sept 2007


22 Sept 2007

  • Transformed into XL-10 gold cells using Quickchange Protocol.

Week 14

23 Sept 2007


24 Sept 2007


25 Sept 2007


26 Sept 2007

  • Miniprepped the following
    • TetR-RBS-P2,21A-ter A,B,C and D
    • TetR-RBS-ComP-ter A,B,C and D
    • GenRBS-ComA-ter A,B,C and D
    • cI-L3 A,B,C and D

27 Sept 2007


28 Sept 2007


29 Sept 2007


Week 15

30 Sept 2007


1 Oct 2007


2 Oct 2007


3 Oct 2007


4 Oct 2007


5 Oct 2007


6 Oct 2007


Week 16

7 Oct 2007


8 Oct 2007


9 Oct 2007


10 Oct 2007


11 Oct 2007


12 Oct 2007


13 Oct 2007