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PCR Analysis of Tail DNA

286 bytes added, 17:00, 8 June 2020

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see [[Genotyping ~~Details~~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 reaction~~

~~Buffer: 4 uL of 5X Go-Taq buffer ("Molecular Biology Stuff" box in freezer)~~

~~Forward Primer: .4ul~~

~~Reverse Primer: .4ul~~

~~dNTPs: .4uL of 2 mM ("Molecular Biology Stuff" box in freezer)~~

~~Sterile water: 13.6 uL~~

~~Polymerase Go-Taq: 1 uL (6th floor in Genotype Yellow Box in freezer)~~

~~Template: 1 uL~~

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

[[Category: Genotyping]]

[[Category: Mouse Work]]

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