Gene polymorphism
Posted by: LLL (IP Logged)
Date: November 30, 2008 03:09PM

Can anyone help me w/my assignment? I've been given part of a gene (240bp; 301-541) with a polymorphism "y" at 405.I must design an assay and describe it in detail (controls, reagents, probes/primers, etc). Any info would be helpful. LL

Re: Gene polymorphism
Posted by: FunctionalBio (IP Logged)
Date: December 04, 2008 12:48AM

Hi, i work for a sequencing service called Functional Biosciences, Inc. We offer SNP resequencing, so I know how to do this if you're going to go with sequencing rather than probes etc.

Just do a PCR reaction, making sure to run negative controls alongside your DNA samples. Do an exo sap clean up of the pcr reaction. Sequence from both directions using ABI big dye v 3.1, just use the pcr primers if the band on the gel looks clean. Theres a program called Mutation Surveyor that will analyze the traces at the polymorphic site. It can objectively designate each sample as a heterozygote or either type of homozygote. We have this software at our lab, its decent. Otherwise i think theres some free snp software here, [snp.wustl.edu]
but i havent used any of it.



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