Neutral polyacrylamide gel electrophoresis.

MATERIALS 5x TBE electrophoresis buffer Polyacrylamide gels are poured and run in 0.5x or 1x TBE at low voltage (1-8 V/cm) to prevent denaturation of small fragments of DNA by Joulic heating. Other electrophoresis buffers such as 1x TAE (please see Agarose Gel Electrophoresis) can be used, but they are not as good as TBE. The gel must be run more slowly in 1x TAE, which does not provide as much buffering capacity as TBE. For electrophoresis runs greater than 8 hours, we recommend that 1x TBE buffer be used to ensure that adequate buffering capacity is available throughout the run. 6x Gel-loading buffer Acrylamide:bisacrylamide (29:1) (% w/v) Ammonium persulfate (10% w/v) Ammonium persulfate is used as a catalyst for the copolymerization of acrylamide and bisacrylamide gels. The polymerization reaction is driven by free radicals that are generated by an oxido-reduction reaction in which a diamine (e.g., TEMED) is used as the adjunct catalyst. DNA samples Ethanol Optional, please see Step 5. KOH/methanol solution Siliconizing fluid (e.g., Sigmacote or Acrylease) (optional) TEMED Neutral Polyacrylamide Gel Electrophoresis -Sambrook and Russell ... file:///C:/Users/MILI%20JOON/Documents/protocol/Electrophoresis,...