I am facing a problem. When I sort the cells by using FACS, I am unable to see CT value in RT PCR. I usually purify more than 10,000 cells and RNA conc also more than 10ng/ul. but when I performed cDNA synthesis, RT PCR then many of the primers have no CT value.
Thanks.
Hi Allah Nawaz ,
I am also dealing with this issue. Did you ever figure out what the problem was?
Hi,
Most importantly to maintain integrity of FACS cells, you need to take precautions during their handling and processing. The RNA extraction process also relates to which type of staining you are doing. Usually you should have higher number of cells for acquisition. So depending on the type of tissues, choose a higher number number (as much possible ) for sorting, so that when you want to extract RNA you have sufficient number of cells.
Also, for reference look this article Article Purification of high-quality RNA from a small number of fluo...
Hope you find some answers.
Best,
Hamid
Please see this article.
Purification of high-quality RNA from a small number of fluorescence activated cell sorted zebrafish cells for RNA sequencing purposes
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