http://2007.igem.org/wiki/index.php?title=Toronto/Lab_Protocols/Overnight&feed=atom&action=historyToronto/Lab Protocols/Overnight - Revision history2024-03-28T21:21:26ZRevision history for this page on the wikiMediaWiki 1.16.5http://2007.igem.org/wiki/index.php?title=Toronto/Lab_Protocols/Overnight&diff=47781&oldid=prevEstherK: /* Miniprep */2007-10-27T03:38:41Z<p><span class="autocomment">Miniprep</span></p>
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<td colspan='2' style="background-color: white; color:black;">← Older revision</td>
<td colspan='2' style="background-color: white; color:black;">Revision as of 03:38, 27 October 2007</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>== <del class="diffchange diffchange-inline">Miniprep ==</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>== Overnight (O/N) Preparation: <ins class="diffchange diffchange-inline">==</ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">Small Scale Plasmid Preparation (1 – 2 hours)</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">'''</del>Overnight (O/N) Preparation:<del class="diffchange diffchange-inline">'''</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Put 2.5 mL of LB Broth into a 15 mL tube. (Make sure to flame the broth)</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Put 2.5 mL of LB Broth into a 15 mL tube. (Make sure to flame the broth)</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Add 2.5 μL of the appropriate antibiotics. (Check spreadsheet)</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Add 2.5 μL of the appropriate antibiotics. (Check spreadsheet)</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Incubate overnight.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Incubate overnight.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"></del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">'''Miniprep Procedure:'''</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"></del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">#Put 1.5 mL O/N into a 1.5 mL Eppendorf tube.</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">#Centrifuge for 1 minute.</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">#Aspirate the supernatant.</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">#In order to increase the concentration of DNA, add the rest of the o/n into the same tube and centrifuge for 1 minute again.</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">#Aspirate the supernatant and resuspend cells with 250 μL Resuspension Solution (red). Vortex until pellet dissolves.</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">#Add 250 μL Lysis Solution and mix with pipette. (shake with hand)</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">#Sit for 3-5 minutes.</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">#Add 350 μL Neutralization Solution (blue) and invert tube with hand 4-6 times.</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">#Centrifuge for 5 minutes.</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">#Pour supernatant in a column tube.</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">#Centrifuge for 1 minute and throw away supernatant.</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">#Add 500 μL Wash Solution.</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">#Centrifuge for 1 minute and throw away supernatant.</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">#Add 500 μL Wash Solution.</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">#Centrifuge for 1 minute and throw away supernatant.</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">#Put blue column in a fresh centrifuge tube. (make sure to label)</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">#Add 50 μL Elution Buffer.</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">#Incubate for 2 minutes at room temperature, then centrifuge for 2 minutes.</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">#Throw away blue column.</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">#Place plasmid into the freezer.</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Jump to [http://igem.skule.ca/lab/protocols/miniprep.htm BlueGenes]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Jump to [http://igem.skule.ca/lab/protocols/miniprep.htm BlueGenes]</div></td></tr>
</table>EstherKhttp://2007.igem.org/wiki/index.php?title=Toronto/Lab_Protocols/Overnight&diff=47770&oldid=prevEstherK: /* Miniprep */2007-10-27T03:37:08Z<p><span class="autocomment">Miniprep</span></p>
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<col class='diff-marker' />
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<td colspan='2' style="background-color: white; color:black;">← Older revision</td>
<td colspan='2' style="background-color: white; color:black;">Revision as of 03:37, 27 October 2007</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>'''Overnight (O/N) Preparation:'''</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>'''Overnight (O/N) Preparation:'''</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">1. </del>Put 2.5 mL of LB Broth into a 15 mL tube. (Make sure to flame the broth)</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Put 2.5 mL of LB Broth into a 15 mL tube. (Make sure to flame the broth)</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">2. </del>Scrape off a single colony from a plate with a pipette tip and eject tip into the tube.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Scrape off a single colony from a plate with a pipette tip and eject tip into the tube.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">3. </del>Add 2.5 μL of the appropriate antibiotics. (Check spreadsheet)</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Add 2.5 μL of the appropriate antibiotics. (Check spreadsheet)</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">4. </del>Incubate overnight.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Incubate overnight.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>'''Miniprep Procedure:'''</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>'''Miniprep Procedure:'''</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">1. </del>Put 1.5 mL O/N into a 1.5 mL Eppendorf tube.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Put 1.5 mL O/N into a 1.5 mL Eppendorf tube.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">2. </del>Centrifuge for 1 minute.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Centrifuge for 1 minute.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">3. </del>Aspirate the supernatant.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Aspirate the supernatant.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">4. </del>In order to increase the concentration of DNA, add the rest of the o/n into the same tube and centrifuge for 1 minute again.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#In order to increase the concentration of DNA, add the rest of the o/n into the same tube and centrifuge for 1 minute again.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">5. </del>Aspirate the supernatant and resuspend cells with 250 μL Resuspension Solution (red). Vortex until pellet dissolves.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Aspirate the supernatant and resuspend cells with 250 μL Resuspension Solution (red). Vortex until pellet dissolves.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">6. </del>Add 250 μL Lysis Solution and mix with pipette. (shake with hand)</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Add 250 μL Lysis Solution and mix with pipette. (shake with hand)</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">7. </del>Sit for 3-5 minutes.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Sit for 3-5 minutes.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">8. </del>Add 350 μL Neutralization Solution (blue) and invert tube with hand 4-6 times.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Add 350 μL Neutralization Solution (blue) and invert tube with hand 4-6 times.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">9. </del>Centrifuge for 5 minutes.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Centrifuge for 5 minutes.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">10. </del>Pour supernatant in a column tube.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Pour supernatant in a column tube.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">11. </del>Centrifuge for 1 minute and throw away supernatant.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Centrifuge for 1 minute and throw away supernatant.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">12. </del>Add 500 μL Wash Solution.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Add 500 μL Wash Solution.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">13. </del>Centrifuge for 1 minute and throw away supernatant.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Centrifuge for 1 minute and throw away supernatant.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">14. </del>Add 500 μL Wash Solution.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Add 500 μL Wash Solution.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">15. </del>Centrifuge for 1 minute and throw away supernatant.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Centrifuge for 1 minute and throw away supernatant.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">16. </del>Put blue column in a fresh centrifuge tube. (make sure to label)</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Put blue column in a fresh centrifuge tube. (make sure to label)</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">17. </del>Add 50 μL Elution Buffer.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Add 50 μL Elution Buffer.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">18. </del>Incubate for 2 minutes at room temperature, then centrifuge for 2 minutes.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Incubate for 2 minutes at room temperature, then centrifuge for 2 minutes.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">19. </del>Throw away blue column.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Throw away blue column.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">20. </del>Place plasmid into the freezer.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Place plasmid into the freezer.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Jump to [http://igem.skule.ca/lab/protocols/miniprep.htm BlueGenes]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Jump to [http://igem.skule.ca/lab/protocols/miniprep.htm BlueGenes]</div></td></tr>
</table>EstherKhttp://2007.igem.org/wiki/index.php?title=Toronto/Lab_Protocols/Overnight&diff=47749&oldid=prevEstherK at 03:34, 27 October 20072007-10-27T03:34:05Z<p></p>
<p><b>New page</b></p><div><br />
== Miniprep ==<br />
<br />
Small Scale Plasmid Preparation (1 – 2 hours)<br />
<br />
'''Overnight (O/N) Preparation:'''<br />
<br />
#1. Put 2.5 mL of LB Broth into a 15 mL tube. (Make sure to flame the broth)<br />
#2. Scrape off a single colony from a plate with a pipette tip and eject tip into the tube.<br />
#3. Add 2.5 μL of the appropriate antibiotics. (Check spreadsheet)<br />
#4. Incubate overnight.<br />
<br />
'''Miniprep Procedure:'''<br />
<br />
#1. Put 1.5 mL O/N into a 1.5 mL Eppendorf tube.<br />
#2. Centrifuge for 1 minute.<br />
#3. Aspirate the supernatant.<br />
#4. In order to increase the concentration of DNA, add the rest of the o/n into the same tube and centrifuge for 1 minute again.<br />
#5. Aspirate the supernatant and resuspend cells with 250 μL Resuspension Solution (red). Vortex until pellet dissolves.<br />
#6. Add 250 μL Lysis Solution and mix with pipette. (shake with hand)<br />
#7. Sit for 3-5 minutes.<br />
#8. Add 350 μL Neutralization Solution (blue) and invert tube with hand 4-6 times.<br />
#9. Centrifuge for 5 minutes.<br />
#10. Pour supernatant in a column tube.<br />
#11. Centrifuge for 1 minute and throw away supernatant.<br />
#12. Add 500 μL Wash Solution.<br />
#13. Centrifuge for 1 minute and throw away supernatant.<br />
#14. Add 500 μL Wash Solution.<br />
#15. Centrifuge for 1 minute and throw away supernatant.<br />
#16. Put blue column in a fresh centrifuge tube. (make sure to label)<br />
#17. Add 50 μL Elution Buffer.<br />
#18. Incubate for 2 minutes at room temperature, then centrifuge for 2 minutes.<br />
#19. Throw away blue column.<br />
#20. Place plasmid into the freezer.<br />
<br />
Jump to [http://igem.skule.ca/lab/protocols/miniprep.htm BlueGenes]</div>EstherK