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Materials
You will need:
- Agarose
- 1X TAE
- Loading Dye
- 2-log or 100bp Ladder
- Safestain
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 a 1:5 volume of loading dye to DNA template. (Eg. 1μL loading dye into a 5μL DNA template to be loaded.)
- Pipette 5μL of ladder into the very first well.
- Carefully pipette each DNA template to each well.
Running the gel
- Run electric current between 90V-120V through gel until bands have separated enough.
Purpose
Although gel electrophoresis has many purposes, we use them to separate out DNA fragments, specifically the insert from the plasmid.
References
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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! :)
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