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-   -   Contamination on PCR real time (http://www.molecularstation.com/forum/real-time-pcr-quantitative-pcr-forum/64940-contamination-pcr-real-time.html)

Lucrecia Carrera 03-03-2009 02:57 PM

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

furqan 03-04-2009 10:14 AM

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 .

Lucrecia Carrera 03-05-2009 09:16 AM

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.

Lucrecia Carrera 03-05-2009 09:16 AM

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.

effendi 03-09-2009 03:00 PM

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


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