Tail DNA for PCR (No Organic Solvents)

1. Obtain the last 2mm of tail and place directly into 200ul 1X PCR Buffer with Nonionic Detergents (PBND) in a 1.5ml microfuge tube. (Tails can be stored at frozen in PBS or PBND until use.)

2. Add 1.2 ul ProK solution (I han't seen any problem with up to 5 ul ProK and tail degredation is slightly faster) to each sample and place in a 55oc bath.

3. Incubate at 55oc with occasional vortexing until tissue is degraded (1-3 hours).

4. Heat samples to 95oc for 10 min. in PCR machine or by boiling to inactivate ProK.

5. Add 2ul(note) processed Tail DNA / 50ul PCR reaction.

note: this volume will vary depending on your particular PCR primers, their Tm etc....

* Adapted from : Perkin Elmer Cetus: Amplifications: Vol. #2 PEC 1989; pp1-3.