Hello everybody,
I am currently extracting total RNA including microRNA from serum using miRNeasy Serum/ Plasma kit Catalog Number 217184 from Qiagen. Within the procedure, we add a Spike -In control to test the recovery of RNA extracted using RT-PCR. The next steps will be library preparation and sequencing using Illumina Machines. Do you expect that the Spike in control will have an effect on the results?
many thanks in advance,
Fawaz
Hi Fawaz Al-Shaheri
since spike-in are well recognized while trimming the raw data of your run, addition of these ones won't change nothing. It depends on the degree of miRNA production of your lineages but it will only take some space in the data range.
fred
You can check these two papers with contradictory results:
Article Assessing the reliability of spike-in normalization for anal...
Article Normalization of RNA-seq data using factor analysis of contr...
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