DNA extraction from mouse ear/tail to genotyping
(NO ORGANIC SOLVENTS EXTRATION)
Obtain the last 2 mm of the ear or tail tissue and place directly into 75 μl Alkaline lysis buffer in a PCR tube. (Tails can be stored at frozen in PBS or PBND until use).
Samples heated in 95oC 10 min –1 h. I actually let them until the tissue is dissolved. After heating, samples are cooled to 4oC and 75 μl Neutralization buffer are added to each sample.
One to five μl of the final preparation are used per each PCR reaction.
Alkaline lysis buffer FINAL VOLUME = 50 ml
25mM NaOH (from 1M)
1/40 –> 1.25ml
0.2mM Na2-EDTA 2H2O (from 1M) (410,31g/mol; 1M = 20.5g in 50ml)
1/5000 –> 10ul
pH: around 12 (not adjust)
Neutralization buffer FINAL VOLUME = 50 ml
40mM Tris-HCl (from 1M) (157,60g/mol; 1M = 7.88g in 50ml)
1/25 –> 2ml
pH: around 5 (not adjust)
Original paper: Biotechniques 2000 Vol 29, No.1 p. 52-54
DNA extraction from mouse ear/tail to genotyping —> PDF