Difference between revisions of "PCR Analysis of Tail DNA"
From Bridges Lab Protocols
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− | see [[Genotyping | + | see [[Genotyping Program]] for strain specific details |
==Materials== | ==Materials== | ||
+ | # Dream Taq Green master mix | ||
+ | # Specific gene Primers (0.4um Working stock) | ||
+ | # Tail digest DNA | ||
+ | # ddH2O | ||
==Protocol== | ==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 | see [[Preparing an Agarose Gel]] for details on preparing a DNA gel | ||
+ | |||
+ | [[Category: Genotyping]] | ||
+ | [[Category: Mouse Work]] |
Latest revision as of 17:00, 8 June 2020
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).
see Preparing an Agarose Gel for details on preparing a DNA gel