mutational studies - (Dec/28/2008 )

Hi,

I'm new to this area of research. If I need to study if a gene is mutated, do I just PCR the whole cDNA, sequence it and check it agaist the pubmed data base? If there any thing else to it?

Thanks

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There's not really much more to it than that, assuming your sequence is solid and the database sequence is solid. But you'll want to compare your sequence to GenBank, not PubMed.

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Hi,

I'm new to mutation studies. My gene of intest has a 1000bp coding region. So, I will design primers from the mRNA seuence and was olanning to sequence the mRNA from cell lines but was told to sequence the genomic DNA to find out if teh mutation in homogenous or heterogenous? What does that mean? Does it mean if the mutation is in either a sngle allelel or both alleles? Also, even if I use the genomic DNA, can I still use the mRNA seq to design my primers?

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But how you will clone both allele. You will use some chromosome marker. Its human gene. I think you have to use the same primer but the may be PCR profile will be different, becasue on genomic DNA there will be lot of introns so gene will be long....

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