Oneill Lab:Gels

Michael J. O'Neill Lab University of Connecticut Department of Molecular and Cell Biology

Agarose gels

 * Electrophoresis grade agarose gels are made with TBE
 * NuSieve gels for DNA purification are made with TAE

Formaldehyde gel
For making a 1% 100 mL
 * 1) Combine
 * 2) *5mL 20x Running Buffer
 * 3) *77mL deionized water
 * 4) *1g agarose
 * 5) Boil in microwave until agarose dissolves
 * 6) Add 18mL 37% formaldehyde
 * 7) *Swirl flask while pouring
 * 8) *Add formaldehyde in fume hood
 * 9) Pour gel in fume hood

Polyacrylamide gels
Add APS and TEMED last. Gel will begin polymerizing immediately.
 * 1) Pour resolving gel first. Be sure to leave space for stacking gel. Will need to allow for length of your comb + 1 cm.
 * 2) Once resolving gel is poured, cover in a layer of iso-butyl alcohol. Leave gel vertical while it polymerizes. This usually takes 45-60 minutes
 * 3) After the gel polymerizes, pour off the alcohol, and rinse the exposed gel with deionized water. Use a paper towel to wick away as much water as possible.
 * 4) Prepare and pour stacking gel to top of plate. Add comb and allow gel to polymerize for 1 hour.

Links and Contacts
Seth Kasowitz [mailto:seth.kasowitz@uconn.edu]

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