DNA extraction from Mouse tails

 Materials :

Lysis buffer
10mM Tris/ HCl
400mM NaCl
2mM EDTA
pH 8,2

Master mix (for one sample)

200µl Lysis buffer
10µl Proteinase K (20U)
50µl 10% SDS

protocol

1. wipe the mouse tail with ethanol and cut the mouse tail from end and obtain  to 2-3mm piece which should be sufficient for the DNA isolation. put it into 1.5 ml eppendroff tube. quickly spin to get all tissue pieces at bottom.

2. Add 260µl of master mix to each sample make sure tail piece is in the solution.

3. Incubate overnight at 56°C in thermal block or waterbath.

Next Day:
4. Heat the sample for 5 min at 96°C.

5. Add 50µl of saturated NaCl and invert the tube 10 times, you will get somemwhat  cloudy solution.

6. Centifuge the tube at max speed (13.2 rpm) for 10 minutes. you will have white pallet.

7. Prepare the another 1.5ml tube with 200µl of isopropanol.

8. Transfer supernatant from step 6 into a new eppendorf tube containing 200µl of isopropanol and invert 25 times. (you can leave the samples at -20 at this stage if you cant complete the protocol today)

9. Spin at max speed for 10 minutes and  Pour off the supernatant  

10.Add 200µl cold 70% EtOH, if pellet moves, spin again Aspirate EtOH and let the pellet air dry 

11.Dissolve in 500µl – 200-µl of TE depending upon size of pallet.

12.Put samples in 50°C for 2 hours before starting the PCR or other recations.

(Note : This protocol can be used for DNA isolation from verious tissues ans cells, for cells you can change incubation to 2 to 4 hours insted of overnight. For tissue isolation from mouse please follow the rules in animal protocol and avoid etical issues. )

 Article By : Team LIFE conjectures