To do liquid biopsy and look for specific gene expressions there are so many published papers.
Thera are different protocols and procedures used to isolate circulating cell free RNA or mRNA from peripheral blood. Some tend to use serum, others used plasma, and some used whole blood.
I have 4 questions:
1- Which one is more efficient to isolate, ccfRNA or ccfmRNA?
2- Do you have any experiments in this field and which one do you used? serum, plasma or whole blood?
3- If you have gone through this, is it efficient to use a simple total RNA isolation kit which can be performed with blood, plasma or serum?
4- To continue with RT-PCR, is it ok to synthesize cDNA from isolated total RNA or mRNA with the commercially available first strand cDNA synthesis kits?
..and continue with qPCR.
https://www.healio.com/hematology-oncology/learn-genomics/introduction-to-liquid-biopsies/what-is-a-liquid-biopsy
Thank you Mr. Srikanth Reddy Madhi Reddy , but my question is about "circulating cell free RNA and mRNA" not DNA.
you can explore two options- exosomal RNA (use Total exosome isolation reagent) and cell free RNA (use MagMax cell-free total nucleic acid isolation kit). They recover different fractions of circulating RNA, and depending on your liquid biopsy "interests" one or another option could be better. I'd recommend to use serum
Serum procedures were conducted on Covid 19 patients here in India they gave excellent results
Thank you Mr. Alexander Sasha Vlassov , Im going to try your suggestions.
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