Team:HKUST/Protocols/PCR cleanup

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 Home Our Team Project Description

<b> Main Parts</li> </b> Odorant Sensing</a></li> Attractant Production</a></li> Toxin Production</a></li>

<b> Resources</li> </b>

Lab Notebook</a></li> Parts Submitted </a></li> Protocol List</a></li> Other Resources</a></li> </ul> <ul> <li><a href="http://2009.igem.org/Team:Gallery">Gallery</a></li> <li><a href="http://2009.igem.org/Team:Biosafety">Biosafety</a></li> <li><a href="http://2009.igem.org/Team:Acknowledgement">Acknowledgement</a></li> </ul> a

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.

<ul> <li><a href="http://2009.igem.org/Team:HKUST/Protocols/Agarose_gel" >Agarose gel preparation and gel electrophoresis</a></li> <li><a href="http://2009.igem.org/Team:HKUST/Protocols/PCR">PCR (Taq, Vent)</a></li> <li><a href="http://2009.igem.org/Team:HKUST/Protocols/PCR_cleanup">PCR product clean-up</a></li> <li><a href="http://2009.igem.org/Team:HKUST/Protocols/Enzyme_digestion"> Enzyme digestion (vector, insert)</a></li> <li><a href="http://2009.igem.org/Team:HKUST/Protocols/plasmid_extraction"> Plasmid DNA extraction</a></li> <li><a href="http://2009.igem.org/Team:HKUST/Protocols/gel_cut"> Gel cutting</a></li> <li><a href="http://2009.igem.org/Team:HKUST/Protocols/gel_extraction"> Gel extraction</a></li> <li><a href="http://2009.igem.org/Team:HKUST/Protocols/Ethanol_precipitation"> Ethanol precipitation</a></li> <li><a href="http://2009.igem.org/Team:HKUST/Protocols/Ligation"> Ligation</a></li> <li><a href="http://2009.igem.org/Team:HKUST/Protocols/Transformation_to_Ecoli"> Transformation to E.coli</a></li> <li><a href="http://2009.igem.org/Team:HKUST/Protocols/Transformation_to_yeast">Transformation to yeast</a></li> <li><a href="http://2009.igem.org/Team:HKUST/Protocols/Cell_lysis"> Cell lysis</a></li> <li><a href="http://2009.igem.org/Team:HKUST/Protocols/Yeast_genomic_DNA_extraction"> Yeast genomic DNA extraction</a></li> <li><a href="http://2009.igem.org/Team:HKUST/Protocols/Western_blotting"> Western blotting</a></li> <li><a href="http://2009.igem.org/Team:HKUST/Protocols/GFP_testing"> GFP testing</a></li>

</ul> iGEM 2009

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