Hiya, I'm hoping someone can help me with an issue I'm having with blood samples on FTA cards.
I have whole blood stored on FTA cards and I want to run ITS2 PCR on the samples.
Current protocol: take punch biopsy, place in pcr tube, add 50 uL NF water, incubate 7 min 53C, quick vortex, pipette supernatant up/down a few times then pipette supernatant out and discard. Repeat once. Then use the washed disc in PCR reaction.
When I do this, the supernatant is not changing colour, becoming pink/orange with the blood cells being washed out of the card disc. The disc is retaining its dark red/brown colour. I worry the retained rbcs interfere with the pcr reaction and could cause false negatives.
I have seen protocols which use an FTA card purification reagent followed by a TE buffer, instead of the NF water. I could try this, but would need to order the products from South Africa (I am based in Malawi).
Does anyone have any advice for me? Thanks a million.
hezy
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