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== | ||
| Line 8: | Line 8: | ||
==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: | Use the following Volumes per 25ul Reaction: | ||
| − | Per sample 1X | + | Per sample (1X) |
#Dream Tag Master mix: 12.5uL ("Molecular Biology Stuff" box in freezer) | #Dream Tag Master mix: 12.5uL ("Molecular Biology Stuff" box in freezer) | ||
| − | #Primer Mix: 5ul | + | #0.4um Primer Mix: 5ul |
#Sterile ddH2O: 7.5ul | #Sterile ddH2O: 7.5ul | ||
| Line 20: | Line 26: | ||
Run "specfic" PCR Program for gene of interest (approx 2 hours). | 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 | ||
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
