Any one working on Porcine endogenous retrovirus (PERV)

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Subramaniam Senthilkumar

Thank u for ur kind reply, we r working on xenograft of porcine origin I just want to quantitate PERV present in the issue sample I am planing to do an infectivity assay by measuring the RT activity using Human embryonic kidney cell line (HEK). I want to know whether any other simple and rapid technique is available to quantitate the virus present in a given sample?

Alessia Armezzani

I guess it depends on what you are looking for...

1) if you want to quantitate PERV viral particles, you may perform a western blot and/or, better, a qRT-PCR;

2) if you want to quantitate INFECTIOUS PERVs, then you have also to perform an infectious assay.

I hope this helps.

Alessia

Thank u for ur comments. I want to quantitate infectious PERV I am planing to do infectivity assay

Hi as u suggested I am doing the infectious assay on HEK cell line by RT activity assay. It has been suggested that we should check the RT activity using the supernatant at an interval of 3 days for 24 days. I would like to know whether i can store the supernatant collected every 3 days and altogether do the RT activity assay on 24th day? or I should do the assay soon after I collected the supernatant?

I am sorry but I cannot help you out with this as I have never performed any RT assay.... I think that, as long as you process the samples in the same way (i.e., you store your last supernatant in the same conditions as the previous ones, let's say for a couple of hours or one day), you should be fine. Alternatively, you could process the supernatant as soon as you collect it, as long as the standard curves for each exp are reproducible...

For the qRT-PCR, I can tell you for sure that it is better to perform the exp as soon as possible and, eventually, store your samples at 4°C, rather than -20°C.

I hope this helps,

Hi I am looking for post doctoral fellowship Is there any position available in your laboratory?

Hi!

I am sorry but in the lab I was for my PhD they have just hired a post-doc and, anyway, they have changed completely subject and are not working on ERVs anymore... if you still want to work on ERVs, you may want to look at lab in London... otherwise, you can try to contact people at the new institute they are building in Glasgow, the Centre for Virus Research (http://www.gla.ac.uk/researchinstitutes/iii/cvr/)... even if at the moment they are officially looking only for a structural biologist, you may find other lab looking for other experties... Good luck! =)

Hi do u ever done the reverse transcriptase activity assay using kit? I am facing some problem.