PCR Analysis of Tail DNA
From Bridges Lab Protocols
see Genotyping Program for strain specific details
Materials
- Dream Taq Green master mix
- Specific gene Primers (0.4um Working stock)
- Tail digest DNA
- ddH2O
Protocol
First, make 250ul of 0.4um working stock primers from 100um Primary Stocks.
- 248 ul ddH20
- 1ul forward primer (100um)
- 1ul reverse primer (100um)
Use the following Volumes per 25ul Reaction:
Per sample (1X)
- Dream Tag Master mix: 12.5uL ("Molecular Biology Stuff" box in freezer)
- 0.4um Primer Mix: 5ul
- Sterile ddH2O: 7.5ul
- Template: 1 uL
Run "specfic" PCR Program for gene of interest (approx 2 hours).
Genotyping Program PCR Amplification of DNA
see Preparing an Agarose Gel for details on preparing a DNA gel