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==EtOH Precipitation== *Pool all reactions into 15ml tube (900ul in total) *add 3ul of glycoblue *add 90ul of 3M NaoAc *add 2250ul of 100% EtOH *Freeze the tube in -80C for 1hr *centrifuge at 3000rpm for 30min, 4C *remove supernatant and add 600ul of chilled 75% ETOH and transfer the DNA pellet to a 1.5ml tube (5000ul-3750ul100% EtOH + 1250ul ddh2o) *spin at 14,000rpm for 5min *dry the pellet in the hood and resuspend w 100ul ddh2o *re-purify the amplicons with 2 Qiaquick columns, elute w 40ul EB for each column *measure concentration w nanodrop
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