Smolke:Protocols/Screening

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Josh's Protocol Sample Preparation Aliquot 50 uL water into sterile PCR tubes, one tube per colony. Using a P20, scape off a colony and resuspend it in the water. Use 1 uL of this cell suspension as your PCR template Reaction Mixture Per colony: 1 uL cell suspension 0.2 uL dNTPs 0.2 uL FWD and REV primers 1 uL 10x Taq Buffer (with MgCl2) 0.2 uL 1:10 Taq 7.2 uL water Note: Make a master mix of everything except the template. Aliquot 9 uL into a separate set of PCR tubes. Then transfer 1 uL of cell suspension to each tube. You can use a multichannel pipet to transfer up to eight templates at once. Run PCR as normal, giving 1 minute/kb for Taq.