I'm trying to amplify an unsequenced region of DNA containing highly repetitive and highly polymorphic genes. I have obtained some good clean sequence but want to expand on this. Does anyone know if it would be possible to obtain small amounts of amplicon if the PCR reaction was done using only one primer? I realise you wouldn't get the exponential growth in product, and the amplicons would be of varying lengths, but if the process was repeated several times and the products purified could you end up with something that could be sequenced?

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hi there,

If I am going to do a PCR amplification of a region in the E. coli genome. I wonder if the first PCR melting temperature should be the same as 95 degree or should it be changed to a much higher temperature? If anyone knows any protocal of doing PCR on genomic DNA plz let me know the links. Thank you!

I am trying to develop a method to synthesize 100-200 bases of ssDNA product from PCR. I have a template lambda DNA. presently using assymetric pcr i am getting more than one strand in denaturng PAGE. To develop my method do i need to use a shorter template as i can reduce the mispriming and unwanted products.
I am using denaturing PAGE gels of 15%, but normal dna ladders 100 bp dont run as i have urea in gel. Can i use a rna ladder.
if have to make this method work with a normal pcr protocol will it be possible. what do i need to do to make my gels work.
I m relatively new to this filed so I am asking this question.
sorry for the long question and thanks in advance for any suggestion.
Regards,
Harish.

I am working in the field of Diabetes genetics. Please let me know which method is best to extract DNA from Blood sample, Here in may lab they have some serum, Plasma and blood cell.it is possible to isolate DNA from them if some one any experience Please share.....