What would be the shortest and optimal method of extracting human cells for PCR? - (Nov/09/2015 )

I am trying to devise a quick method to extract genomic DNA from human cells for a PCR.

I first collected cells by centrifuging saline mouthwash (0.9% NaCl) and extracted genomic DNA using kit followed by a PCR protocol which works fine but is a rather lengthy process.

I tried to cut short the method and removed the DNA extraction steps by using pelleted spit samples as templates for PCR and increased the initial denaturing step from 3 minutes to 5 minutes. However this method was not reliable as some of my samples were amplified and some were not.

Can anyone suggest me a reliable protocol here?

Thanks!

-techtrix-

If you have too much template it may inhibit the pcr. You can try pellet the cells, wash with pbs 1x, boil 5 min more or less and use this as template for pcr. Before using in the sample make a test with your own sample and other person that can contribute with sample.

-merlav-

The most common problem is having too much template (with inhibitors). Less is more. Dilute and use small amounts.

-phage434-