Mouse tissue lysis for genotyping protocol
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- Taq DNA polymerase 10X buffer with MgCl2 (diluted 1:10 in ddH2)
- proteinase K (20 mg/ml)
- tail or tissue chunk
- Tissue lysis to release DNA
Measure out 100 µl of Taq DNA polymerase 10X buffer with MgCl2 into tail or tissue chunk.
Add 2 µl of proteinase K.
- Incubate at 50°C for 3 - 12 hrs(overnight) with shaking at 300 rpm.
proteinase K is stable over a broad pH range (4.0 to 12.5, optimum pH 8.0) and is also stable over the temperature range of 25 to 65°C.
- Proteinase K inactivation
Incubate at 95°C for 10 - 20 mins with shaking at 300 rpm.
Centrifuge at a speed of 13000 rpm for 3 mins at room temperature and aspirate out 80 µl of top layer into sterile 1.5-ml microcentrifuge tube (2).
Discard bottom layer.
- Storage before PCR
NOTE: If sample is/are used within 1 hr, set temperature to 4°C and use, else store it at -20°C.