SEVERE PROBLEMS WITH BUCCAL SAMPLES - (Feb/12/2008 )
I used the BuccalAmp™ DNA Extraction Kit, Catch-All™ Sample Collection Swabs, to collect 100 samples for lct gene islolation and amplification. When I did the PCR last november i was getting fairly good results. Even a month ago my control sample (which used the same kit) was working, although not too well. However, none of my samples seem to be working now. i've tried changing temperature, water, salt content, new enzymes. And i'm still getting bupkis... Has anyone used this kit before? and would anyone have any possible ideas what could have gone wrong?
I had used the buccal amp and you will not get tons of DNA, but there are some tips to get enough sample for the PCRs
You don't say what type of sample, so I will think that you work with human subjects.
1. Coffee is a PCR "killer" so no coffee at least 30 min before the sample taking.
2. subjects must rinse with plain water 2-3 times.
3. subject must pass the swab 30-40 times over the gums and cheeks (never the teeth).
4. When extract the cell from the swab place the swab in the buffer, cut the the handle and vortex a lot, with a tweezer take out the swab and press against the walls of tube to get the the excess of liquid and keep the swab in another tube. Just in case!!!
5. follow the protocol as the kit say.
thanks, I should be recieving some new samples soon, so i'll take your advice. Unfortuneatly i've now got to try and trouble shoot to get the already extracted samples to work.