Mouse tissue lysis for genotyping

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(*** page creation *** lead section)
(lysis steps)
Line 1: Line 1:
This is a quick protocol for mouse tail and tissue lysis with proteinase K. It is commonly used to prepare templates for genotyping. Other protocols included detergents in the lysis buffer, but we found this simple protocol to work well with less hands-on time.
This is a quick protocol for mouse tail and tissue lysis with proteinase K. It is commonly used to prepare templates for genotyping. Other protocols included detergents in the lysis buffer, but we found this simple protocol to work well with less hands-on time.
 +
 +
tissue lysis to release DNA:
 +
* per tail or tissue chunk (tissue degradation is sped up if tissue/tail is cut into smaller bits):
 +
** 100 µl Taq DNA polymerase 10x Reaction buffer with MgCl2 (e.g. Promega #M1883) diluted 1:10 in ddH2O
 +
** +2 µl proteinase K (20 mg/ml)
 +
* incubate for 3h to o/n at 56°C at 300 rpm

Revision as of 09:05, 18 April 2007

This is a quick protocol for mouse tail and tissue lysis with proteinase K. It is commonly used to prepare templates for genotyping. Other protocols included detergents in the lysis buffer, but we found this simple protocol to work well with less hands-on time.

tissue lysis to release DNA:

  • per tail or tissue chunk (tissue degradation is sped up if tissue/tail is cut into smaller bits):
    • 100 µl Taq DNA polymerase 10x Reaction buffer with MgCl2 (e.g. Promega #M1883) diluted 1:10 in ddH2O
    • +2 µl proteinase K (20 mg/ml)
  • incubate for 3h to o/n at 56°C at 300 rpm
Personal tools