Post Electrophoretic Analysis Articles
Gel Preparation for Native PAGE of DNA

An exploded view of a gel cassette.
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.
The table below gives solution amounts to prepare gels of various percentages and buffers.
Instructions:
- Prepare a solution of monomer, buffer, and water as per the table.
 - De-gas if desired.
 - Add APS and TEMED.
 - Pour gel.
 
| 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  | 
NEXT TOPIC: PCR Analysis: An Examination
- Using PAGE to Determine Nucleic Acid Molecular Weight
 - SSCP Analysis
 - Sanger Sequencing
 - Sample Preparation for Native PAGE of DNA
 - Sample Prep for Denaturing PAGE of DNA
 - S1 Mapping
 - Run Conditions in Denaturing PAGE
 - RNA Mapping
 - RNA Electrophoresis
 - Ribonuclease Protection
 - Restriction Digest Mapping
 - Primer Extension
 - Preparing Denaturing DNA & RNA Gels
 - Preparation of Denaturing Agarose Gels
 - Preparation of Agarose Gels
 - Pouring Sequencing Gels
 - PFGE and FIGE
 - PCR Analysis: Yield and Kinetics
 - PCR Analysis: An Examination
 - Native PAGE of DNA
 - Mobility Shift Assay
 - Methylation & Uracil Interference Assays
 - Maxam & Gilbert Sequencing
 - Manual Sequencing
 - In Gel Enzyme Reactions
 - Heteroduplex Analysis
 - Gel Preparation for Native PAGE of DNA
 - Gel Electrophoresis of PCR Products
 - DNase I Footprinting
 - DNA/RNA Purification from PAGE Gels
 - DNA/RNA Purification from Agarose Gels – Electroelution
 - Differential Display
 - Denaturing Polyacrylamide Gel Electrophoresis of DNA & RNA
 - Conformational Analysis
 - Automated Sequencers
 - Analysis of DNA/Protein Interactions
 - Agarose Gel Electrophoresis of DNA and RNA – Uses and Variations
 - Agarose Gel Electrophoresis of DNA and RNA – An Introduction
 
