Genomic DNA Extraction from K. marxianus

Amresco yeast genomic DNA extraction Kit:

  1. Centrifuge 1ml of cells at 1000 g for 2 minutes. Remove the supernatant.
  2. Add 300μl of Yeast Lysis solution and 10μl of RNase A provided with the kit. Resuspend the pellet by vortexing and incubate at 65°C for 15 minutes then chill the samples on ice for 8 minutes.
  3. Add 100μl of protein precipitation solution provided by the kit. Vortex for 15 seconds.
  4. Centrifuge at max rpm for 15 minutes.
  5. Transfer the supernatant to a clean 1.5ml centrifuge tube.
  6. Add 500μl of isoproponal and mix via inverting the tube 5-10 times.
  7. Centrifuge again for 10 min at max rpm to pellet the DNA. Carefully discard the supernatant.
  8. Add 500μl of 70% ethanol to wash the pellet.
  9. Centrifuge at max rpm for 2 minutes and carefully discard the supernatant.
  10. Air dry the pellet for 10 minutes.
  11. Resuspend the genomic DNA in 40μl TE Buffer.