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Team:HKUST/Protocols/PCR cleanup
From 2009.igem.org
3. PCR product clean-up
Purpose: To clean up primers, dNTPs, enzymes, short-failed PCR products in the PCR reaction. Materials: FAPC Buffer，Wash Buffer, Elution Buffer, PCR Product (all provided in FavorPrepTM PCR Clean-Up Kit) Procedure: a.Transfer 30μL of PCR product and add 5 volumes of FAPC buffer to a 1.5mL microcentrifuge tube, then mix well by vortexing. b.Place a FAPC Column into a Collection Tube and transfer the sample mixture to FAPC Column. c.Centrifuge at 6,000 rpm for1 min then discard the flow-through. d.Add 750 μl of Wash Buffer (ethanol added) to FAPC Column. Centrifuge at 6,000 rpm for 1 min, then discard the flow-through. e.Centrifuge at 14,000 rpm for an additional 3 min to dry the column. f.Place FAPC Column into an Elution Tube. g.Add 40 μl of Elution Buffer or ddH2O (pH 7.0~8.5) to the membrane center of FAPC Column. Stand FAPC Column for 2 minutes. h.Centrifuge at 14,000 rpm for 1 min to elute the DNA. Tips: •In step f, for effective elution, make sure that the elution solution is dispensed onto the membrane center and is absorbed completely.
