I have obtained 100 ul of pre-amplified cDNA using the SSIV SINGLE CELL PRE-AMP KIT. Now i want to make a dilution because my samples are limited and i cannot directly use the cDNA. So what dilution should i use and decide on for the cDNA template when using the Applied Biosystems™ TaqMan® Gene Expression Master Mix (Cat. No. 4369016).
If c-DNA is synthesized from 2 micro gram RNA, you can take 1:10 dilution of that c-DNA .
1) why are you running qPCR on a scRNAseq library? This seems like an extravagant waste of incredibly expensive sample material.
2) you specify PRE AMP: have you considered amplifying the library first?
3) 1/10 to 1/20 should be fine, depending on your target. If you've got 100ul, take 5ul and dilute it into 45ul H2O, then run a couple of qPCRs to get a feel for your target Cq. If it's
You can make a dilution curve to select the best dilution. Generally, 1:10 or 1:20 works well, however, it depends on your target genes.
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