Genotyping- PCR
PCR- 2% Agarose Gel Preparation
1. Assemble gel chamber and ensure that there is a tight seal (agarose solution is
poured within the mold)
2. Insert combs to produce wells
3. Prepare 1000ml of 1X TAE Buffer
[980ml ddH2O/ milli-Q + 20ml 50X2 TAE]
4. Take 100mL of 1X TAE
5. Add 2g of Agarose powder1 to the 100ml of 1x TAE Buffer
6. Mix the solution and Microwave until the solution is transparent and
homogeneous
7. Add 10 µL of Syber safe3 to the solution and mix well
8. Pour the gel solution into the chamber avoiding air bubbles
9. Let solution polymerize for ~20mins
10. Add 800-900ml of 1x TAE buffer into gel chamber
11. Then remove combs (combs cannot be removed before adding the buffer because
the gel would be damaged in the process)
1
UltraPure Agarose, #16500-500 Company: Invitrogen
50X TAE Buffer #B49 Fisher
3
SYBRsafe DNA gel stain 41105321
2
Updated by LM on 4/21/15
1