Gel Preparation for Native PAGE of DNA

Native PAGE gels are prepared by mixing an acrylamide/bisacrylamide monomer concentrate (AccuGel 19:1 or 29:1), buffer concentrate and water to achieve the desired gel concentration. TEMED and ammonium persulfate are added to initiate polymerization and the solution is poured into a cassette. The comb is then inserted.

The cassette is formed by two glass plates separated by spacers, typically 0.5-1mm in thickness, and sealed at the bottom (See figure below or follow this link for a more detailed discussion of the electrophoresis apparatus). The polymerization process takes 1-2 hours to complete.

Gel Cassette
An exploded view of a gel cassette.

 

The table below gives solution amounts to prepare gels of various percentages and buffers.

Instructions:

  1. Prepare solution of monomer, buffer and water as per table.
  2. De-gas if desired.
  3. Add APS and TEMED.
  4. Pour gel.

Formulas for Native DNA Polyacrylamide Gels using National Diagnostics' AccuGel 40%

Gel % TBE Gels TAE Gels TTE Gels
6 AccuGel: 15 mL
10X TBE: 10 mL
Deionized H2O: 75 mL
AccuGel: 15 mL
50X TAE: 2 mL
Deionized H2O: 83 mL
AccuGel: 15 mL
20X TTE: 5 mL
Deionized H2O: 80 mL
8 AccuGel: 20 mL
10X TBE: 10 mL
Deionized H2O: 70 mL
AccuGel: 20 mL
50X TAE: 2 mL
Deionized H2O: 87 mL
AccuGel: 20 mL
20X TTE: 5 mL
Deionized H2O: 75 mL
10 AccuGel: 25 mL
10X TBE: 10 mL
Deionized H2O: 65 mL
AccuGel: 25 mL
50X TAE: 2 mL
Deionized H2O: 78 mL
AccuGel: 25 mL
20X TTE: 5 mL
Deionized H2O: 70 mL
12 AccuGel: 30 mL
10X TBE: 10 mL
Deionized H2O: 60 mL
AccuGel: 30 mL
50X TAE: 2 mL
Deionized H2O: 73 mL
AccuGel: 30 mL
20X TTE: 5 mL
Deionized H2O: 65 mL
14 AccuGel: 35 mL
10X TBE: 10 mL
Deionized H2O: 55 mL
AccuGel: 35 mL
50X TAE: 2 mL
Deionized H2O: 68 mL
AccuGel: 35 mL
20X TTE: 5 mL
Deionized H2O: 60 mL


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