Hi everybody!
I'd like to generate and quantify whole transcriptomes by using RNA-seq in the near -- well, not so near -- future but I am completely newbie on this method.
My doubt concerns mainly the quantification capacity of RNA-seq: how can RNA-seq be used to quantify transcripts if a PCR amplification has to be done after the adapter ligation in the library preparation setp? How can this amplification step not impair the post-sequencing quantification of transcripts?
Is the amount of PCR-amplified products directly proportional to the initial amount of cDNAs in this case?
Thank you in advance!
Marcio
I'd like to generate and quantify whole transcriptomes by using RNA-seq in the near -- well, not so near -- future but I am completely newbie on this method.
My doubt concerns mainly the quantification capacity of RNA-seq: how can RNA-seq be used to quantify transcripts if a PCR amplification has to be done after the adapter ligation in the library preparation setp? How can this amplification step not impair the post-sequencing quantification of transcripts?
Is the amount of PCR-amplified products directly proportional to the initial amount of cDNAs in this case?
Thank you in advance!
Marcio
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