Lab 3.4 2012
Lab 3.4 2012 kdorfman Tue, 03/26/2013 - 15:55M 3/26/12
CORRECT THE VOLUMES IN THE LAB MANUAL 21 mL 10X in 210 mL total!
Goal for this lab: Genotype Salk line plants, looking for homozygous mutants.
Use 3 primers in a single reaction (GSP1, GSP2, LBP)
LB 195 LBb1.3 ATTTTGCCGATTTCGGAAC (111 bp from left border)
Materials
- PCR strip tubes
- Harris cutting mat
- Harris Uni-Core (0.5 mm) Cutting Tool
- Beakers for bleach
Reagents
- 2% bleach (12 mL bleach + 588 mL dH2O)
- 5U/µL Taq (3.5 µL per pair)
- 10X polymerase buffer (21 + 21 = 42 µL per pair)
- 2.5 mM dNTP mix (51 µL per pair)
- Sterile dH2O
- Working stocks of primers, including LBP diluted to 10 µM (17µL/rxn)