Contamination on PCR real time
 

Hi

I hope someone can help me whit this.

Im doing a research in humans so im using a sample of lymphocytes and im doingo real time PCR. Using a GRS primers and 36B4Ribosomal as a housekeeping.
I got contamination, my negative amplifies and my syber green, the water and the primers got contamination every time. I change everything all the time but im tired to got this so often. So i decided to work on the laminar flow cabinet. And a got contamination so i dont whats happening and dont know what else to do.

Can any body help me? some ideas??? i try almost everything :(

Thanks

Lu

Re: Contamination on PCR real time
 

furqan: thnk your pippetes set had contaminated try to change them & always used autoclaved tips, eppendorf tubes & sterile conditions, hopefully this will work for u .

Re: Contamination on PCR real time
 

Thanks... i will try to change the pippetes because all the other things i used are autoclaved always...
Tell u something thank u.

Re: Contamination on PCR real time
 

Thanks... i will try to change the pippetes because all the other things i used are autoclaved always...
Tell u something thank u.

Re: Contamination on PCR real time
 

Try to use micropippete tips which have a filter. Wash your micropippete with 10% sodium chloride (wipe the micropippete with NA chloride) before used and keep all stuffs (except DNA/RNA/cDNA) on the bench (laminar flow) with uv light for 15 min or more.
hope it works