We know already that the Kapa PCR Blood kit can work with whole blood as a template in order to do PCR directly and many of you have asked me about the possibility to do the same with mouse tails (and of course with other samples like plant tissue, cultured cells, sperms etc.).

Today we would like to show you a very promising combination of Nippon Genetics DNAreleasy kit and the new Kapa 2G Robust HS enzyme.

Everybody knows that extraction of genomic DNA from mouse tails and tissue samples is a very time consuming procedure. Many protocols in the WEB describing an overnight incubation of mouse tails with proteinase K because the cell lysis step is very difficult.

The best procedure until now, is to use a spin column kit and do a hot start PCR after the purification. This is not as time consuming as the traditional method but of course it is more expensive.

A very powerful method, shown here is the combination of DNAreleasy and the Kapa 2G Robust HS enzyme:

Competitor V (overnight incubation) Pro K digestion (overnight incubation)

DNAreleasy (7 min. incubation) Homemade method (10 min. incubation)

To a 2 mm segment of mouse tail, 50 µL of DNAreleasy (Nippon Genetics Europe) was added.
Samples were incubated for 5 minutes at 75ºC, followed by 2 minutes at 96ºC. After lysis, 2 µL of product was used in 25 µL PCR reactions of 8 amplicons using KAPA2G Robust HotStart ReadyMix.