postivie control of HIV-1 Amplicor

Dr. J.P. Clewley jclewley at hgmp.mrc.ac.uk
Fri Aug 30 03:15:44 EST 1996


Alex Chang writes:

>
>Out of curiosity, I did an electrophoresis of the positive control (from
Amplicor HIV-1 Amplification Kit) PCR amplified using unlabelled primers,
SK462 and SK431.

To my surprise, the amplified product is only around 100 bp. In my
impression, the positive control product should be the same length (142
bp) with the target, HIV-1 gag. 


I reply:

You may be interested in a paper in J Virol Methods by Barlow et al
52:65-74, 1994 where the positive control is also run on a gel and
appears to be the correct size of 142 bp. There is a lower MW shadow
band tho'.

Did you run clinicals alongside the pos. control?
How did you renature the amplicon from the alkali?

I don't know for sure what the control is but I assume its a plasmid
as the protocol booklet with the kit says that the sensitivity was
determined with a plasmid titration series.

I'd also be interested in any other details/experience with this
kit.

Jon Clewley



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