PCR Analysis of Tail DNA

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see Genotyping Details for strain specific details

Materials

  1. Dream Taq Green master mix
  2. Specific gene Primers (0.4um Working stock)
  3. Tail digest DNA
  4. ddH2O

Protocol

First, make 250ul of 0.4um working stock primers from 100um Primary Stocks.

  1. 248 ul ddH20
  2. 1ul forward primer (100um)
  3. 1ul reverse primer (100um)


Use the following Volumes per 25ul Reaction:

Per sample (1X)

  1. Dream Tag Master mix: 12.5uL ("Molecular Biology Stuff" box in freezer)
  2. 0.4um Primer Mix: 5ul
  3. 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