Mouse Tail DNA Analysis Protocol

Overview

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Procedure

Conklin Lab Tail Tip Digest Protocol

(Modified protocols from Hanahan Laboratory at UCSF)

  1. Cut 1/16" to 1/8" of mouse tail from mouse using a razor blade.
  2. Place tails in labeled 1.5 ml eppendorf tubes.
  3. Prepare digest mixture:
     Add 17 microliters tail tip buffer and 3 microliters proteinase K (20 mg/ml) for a total of 20 microliters
  4. Add 20 microliters of digest mixture into tubes containing tail tip.
  5. Incubate tubes in a 55 deg C water bath for 1-2 hours.
  6. Remove tubes from water bath. Spin briefly for 10 seconds.
  7. Add 500 ul of milli-Q water to each tube.
  8. Boil samples for 5 minutes. Place weight on top of tubes to prevent caps from popping.
  9. Spin samples briefly. Store at 4 deg C until PCR. Tail tip buffer: 50 mM Tris, pH 8.0 20 mM NaCl 1 mM EDTA, pH 8.0 1% SDS (adjust to proper volume using milli-Q water) 

Conklin Lab PCR Tail Tip Analysis Protocol

3. Prepare PCR reaction mixture:

   39.8 microliters Milli-Q water 5.0 microliters 10x PCR buffer +Mg 1.0 microliter 10mM dNTP 1.0 microliter 5' primer (25uM) 1.0 microliter 3' primer (25uM) 0.2 microliter Taq (added last) Total = 48.0 microliters

(Taq and 10x PCR buffer +Mg are from Boehringer Mannheim)

   $ 6. Analyze PCR product on a 2% agarose gel. : ==References==

Relevant papers and books

If this protocol has papers or books associated with it, list those references here. Below is an example for formatting purposes. See the OpenWetWare:Biblio page for more information.

1. Goldbeter A and Koshland DE Jr. An amplified sensitivity arising from covalent modification in biological systems. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6840-4. DOI:10.1073/pnas.78.11.6840 |
2. JACOB F and MONOD J. Genetic regulatory mechanisms in the synthesis of proteins. J Mol Biol. 1961 Jun;3:318-56. DOI:10.1016/s0022-2836(61)80072-7 |
3. ISBN:0879697164

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