Question:
What is the process of RNA sequencing?
Author: Mirte SchattorieAnswer:
1. RNA isolation from sample. 2. isolate specific RNA species you want to analyse, e.g. isolate miRNAs through size selection or other RNAs through poly-A selection. 3. conversion into cDNA. 4. add adaptors to both ends of cDNA fragments. 5. align the fragments with the reference genome. 6. classify reads as either exonic reads, junction reads and poly(A) end-reads. 7. the result is quantified by counting the number of reads that align to each gene using a computer program.
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