I wanted to detect HPV in Human DNA samples using touch down PCR as I believe that Touch down is a more sensitive to low copy number of viral DNA in trying to detect HPV in human DNA.
Personally, I have made a set of primer that detects several strains of HPV which works great with high sensitivity. However, to really determine that there are really NO HPV DNA in the human DNA. I used a general primer system GP5+ and GP6+ which is used widely across various labs for detection of HPV.
Apparently, when I used the GP5/6 system. I had very random results. Some of the samples showed 100% positive. Some showed 0%. The worst thing is, under the system my B-actin gets amplified 50% of the time.
I had no problems with amplifying B-actin in my own detection system which also uses Touch down.
What is the problem with GP5+/6+ system.
Both are positive for HPV 18.
Lane 1 - DNA marker
Lane 2 - Hela cell DNA (GP system)
Lane 3 - Cervix cells DNA (GP system)
Lane 4 - Hela cell DNA (B-actin)
Lane 5 - Cervix cells DNA (B-actin)
Lane 6 - Hela cell DNA (my system)
Lane 7 - Cervix cells DNA (my system)
Lane 8 - Hela cell DNA (B-actin)
Lane 9 - Cervix cells DNA (B-actin)
Touch down PCR problem
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