Team:UC Davis/protocols/gelelectrophoresis.html

From 2010.igem.org

(Difference between revisions)
m
m
Line 27: Line 27:
                 <li>Agarose </li>
                 <li>Agarose </li>
                 <li>1X TAE </li>
                 <li>1X TAE </li>
 +
                <li>Loading Dye</li>
 +
                <li>2-log or 100bp Ladder </li>
                 </ul><p>
                 </ul><p>
Line 59: Line 61:
<ul>
<ul>
<li>Pipette 1μL of loading dye for every 5μL of DNA template</li>
<li>Pipette 1μL of loading dye for every 5μL of DNA template</li>
 +
<li>Pipette 5μL of ladder in the very first well. </li>
<li>Carefully pipette each DNA template to each well.</li>
<li>Carefully pipette each DNA template to each well.</li>
</ul>
</ul>

Revision as of 21:59, 9 September 2010

Gel Electrophoresis

Materials

You will need:

  • Agarose
  • 1X TAE
  • Loading Dye
  • 2-log or 100bp Ladder

Extra Notes

  • Be sure that the well size is large enough (but not too large) to accommodate the amount of DNA you will be loading.
  • Be sure to aware of what concentration your gel is. The difference between a 1% and a 2% agarose gel is significant.

Procedure

 Creating a 1% gel stock
  • Dissolve 5g agarose in 500mL 1X TAE

Creating a 2% gel stock

  • Dissolve 10g agarose in 500mL 1X TAE

When gel is ready to use

  • Have gel trays ready with the gel comb (used to create the wells) in place.
  • Microwave the gel stock. Be sure it is all melted and dissolved.
  • Pour 50mL into one 50mL centrifuge tube. If gel is bigger, pour 100mL into two 50mL centrifuge tubes.
  • When gel solution is lukewarm, pipette 5μL safestain into gel
  • Pour immediately and slowly into gel trays to reduce chunks and bubbles in gel.
  • Wait until gel completely solidifies.

Loading DNA into gel

  • Pipette 1μL of loading dye for every 5μL of DNA template
  • Pipette 5μL of ladder in the very first well.
  • Carefully pipette each DNA template to each well.

Purpose

 Although gel electrophoresis has many purposes, we use them to separate out DNA fragments, specifically the insert from the plasmid.

References
We would like to take a moment to thank all of our sponsors for their very generous donations, as we could not have done this without your help!

We would also like to thank and acknowledge:
Our Advisors
Marc Facciotti
Ilias Tagkopoulos
Technical Guidance
David Larsen
Andrew Yao
Visiting iGEMer
Jia Li of Zhejiang University (TEAM ZJU-China)
cI Promoter Screen
Drew Endy - Stanford
Thomas Schneider - NIH
Want to sponsor us? Send an email to mtfacciotti@ucdavis.edu to discuss various ways you can help! :)

Retrieved from "http://2010.igem.org/Team:UC_Davis/protocols/gelelectrophoresis.html"