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==EtOH Precipitation== *add 1ul of glycoblue *add 10ul of 3M NaoAc *add 250ul of 100% EtOH *Freeze the tube in -80C for 30min *centrifuge at 10000rpm for 20min, 4C *no pellet seen after centrifuge, added 1ul extra glycoblue and freeze for 1 hour, will spin at 14000 rpm for 30 min *doble the volume of all etoh precipitation reagents and spin again, this time we are able to see a blue pellet *remove supernatant and add 500ul of chilled 75% ETOH and transfer the DNA pellet to a 1.5ml tube (5000ul-3750ul100% EtOH + 1250ul ddh2o) *spin at 10,000rpm for 5min *dry the pellet in the hood and resuspend w 100ul ddh2o
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