Extraction of chromosomal DNA with PCA (phenol:chloroform:isoamylalcohol) - MetabolicEngineeringGroupCBMA/MetabolicEngineeringGroupCBMA.github.io GitHub Wiki

This protocol is quick but also dirty. There should be considerable amounts of phenol in the final preparation. Do not use for E. coli transformation, the phenol kills the E.coli.

  • Glass beads (~150 µL)

  • 150 µL TE 10 X

  • 150 µL PCA (phenol : chloroform : isoamyl alcohol at proportion 25:24:1) (Plant lab)

  • 20 sec Fast Prep (or 5 min; max rpm; in the LGM disruptor)

  • Centrifuge 10 min 14 000 rpm

  • DNA is in the upper phase

  • Dilute 10x and check in the NANODROP

  • Use it as a template.

Reference for this protocol?

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