Hello everyone please i wish to understand why cDNA is done on RNA samples before RNA sequencing is done. is it possible to just construct the library directly from the RNA sample without first doing cDNA prior to library construction?
The conversion of RNA to cDNA before RNA sequencing is a common and well-established practice because it provides a stable and amplifiable form of the RNA for library construction, offers the ability to selectively capture mRNA, and helps overcome some of the challenges associated with RNA stability and degradation.
Hello Alex, The conversion of cDNA from RNA facilitates the stability and PCR amplification for the template. Its usually not recommended (Takes a lot of input material due to no amplification). However if you want to perform direct RNA sequencing there are options available with Oxford Nanopore with option RT (you can look it up)
Can you tell me which RNA you are trying to sequence like is it Total RNA, mRNA or some other type with high methylations like tRNAs or rRNAs.
Hello Ambika Dudhate i'm trying to sequence Total RNA from human stool samples basically the downstream of my DNA/RNA extraction is for viral metagenomics. i'm using the Purelink viral DNA/RNA kit and the company says i need to pay extra for cDNA to be done on the extracted RNA sample prior to RNA seq.
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