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L A B O R A T O R Y
Part III: DNA Analysis by Gel Electrophoresis

Reagents Equipment & Supplies Shared Items
2% agarose gel
1X electrophoresis buffer
pBR322-BstNI markers
1 µg/ml ethidium bromide or 0.05% methylene blue staining solution
15 ml test tube, polypropylene
1-20 µl micropipet and tips
Staining tray or weigh boat 		Electrophoresis chamber
Electrophoresis power supply

Procedure

  1. Use a micropipet with a fresh tip to add entire PCR sample/loading dye mixture into your assigned well of a 2% agarose gel. Expel any air from the tip before loading, and be careful not to push the tip of the pipet through the bottom of the sample well.
  2. Load 5 µl of the pBR322-BstNI size markers into one lane of gel.
  3. Electrophorese at 130 volts for 20-30 minutes. Adequate separation will have occurred when the cresol red dye front has moved at least 50 mm from the wells.
  4. Gel may be stained either with 1 µg/ml ethidium bromide for 10 minutes or 0.05% methylene blue (or proprietary) stain for 30 minutes, followed by 20-30 minutes destaining with water.
A L U  I N S E R T I O N  P O L Y M O R P H I S M

DNA Learning Center, Cold Spring Harbor Laboratory
Noncommercial, educational use only.
Kits available from Carolina Biological Supply Company.