Protocols

 

DNA Extraction for ES-Cell in 96-well Plates

1. Remove media with multiple chanel pipette or by aspiration.
2. Add 100 µl PBS, remove
3. Add 50 µl lysis buffer
4. Wrap the plate with wet paper towel then seal it in a plastic bag
5. Incubate at 50°C overnight
6. Add 100 µl cold 100% etOH
7. Leave at room temp (not shaking)
8. If a plate spinner is available, spin plates for 5’ at 1000 rpm at 10°C. Pour off EtOH by blotting on paper towel pad.
9. Wash 3 x with 150 µl 70% EtOH and pour off
10. Pat on paper towel pad until very little liquid comes out
11. Dissolve in 25 µl 0.1 x TE or 50 µl Tris 10 mM for PCR , incubate the plate at 55°C for one hour in a SEALED bag or Saran Wrap with wet towels. After dissolving, store the plate at 4°C or do PCR screening directly.
12. Digest the DNA in 30 µl total volume overnight at 37°C.

Lysis Buffer

1 mg/ml proteinase K