If I am interested in a customized mRNA-only Sequencing (RNA-Seq) based on few transcripts (not the whole transcriptome), and after having satisfied & permissible RIN values for my extracted RNA, if now I only perform the reverse transcription through highly efficient cDNA approaches like using SSIV VILO mastermix with oligo DTs priming to enhance my mRNA only-yield by eliminating the random hexamers at all in my reaction to rule out any possibility of rRNA conversion.
So, would it be suffice or I must need to avail rRNA removal kits like RiboMinus, RiboErase, Clean NGS etc.
If there is no choice except to use such kits and protocols, so which one is more efficient to do so? Any experiences or ideas !!!!!
If you use oligo-dT, you don't need to perform rRNA depletion. That's what we usually do for total mRNA sequencing and it works quite well. Now, if you're interested only in a few transcripts, is it not possible to reverse transcript only these mRNA so to have a better sequencing depth and to be able to multiplex your samples at a larger scale?
- Badges
- Science method