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.
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| An exploded view of a gel cassette. |
The table below gives solution amounts to prepare gels of various percentages and buffers.
Instructions:
- Prepare solution of monomer, buffer and water as per table.
- De-gas if desired.
- Add APS and TEMED.
- Pour gel.
Formulas for Native DNA Polyacrylamide Gels using National Diagnostics' AccuGel 40% |
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| 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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