Team:Aachen/Wetlab/Protocol19

Genome Isolation for S. cerevisiae:

Procedure:


  1. pick a large colony and resuspend in 100 uL 0.2 M LiOAC containing 1% SDS.

    • Solution: mix 1,3196 mg LiOAC, 0,67 mg SDS and 100 µl ddH2O

  2. incubate 10 min at 75 ℃.

  3. add 300 µL 100 % EtOH, vortex.

  4. centrifuge 3 min at max. rpm, discard supernatant.

  5. resuspend pellet in 150 uL 70 % EtOH (→ for 70% EtOH mix 105 ul of 100%-EtOH with 45 ul H2O).

  6. centrifuge 3 min at max. rpm, discard supernatant.

  7. dry pellet at 37 ℃ (place tube upside down).

  8. add 50 uL ddH2O to the dry pellet and vortex shortly.

  9. centrifuge 1 min at max. rpm, use 2 uL supernatant for PCR.