We need to make Illumina libraries from small quantities (<1ng) of human mRNA than has been degraded to a fragment size <150 nt.
We can do this by randomly-primed RT, second strand synth, end prep and adaptor ligation, a la standard RNASeq.
Or we can do it by sequential ssRNA adaptor ligation, a la miRNASeq.
Which would people advise using?
In favor of the standard approach is that we do this method all the time.
In favor of the miRNA approach is that it gives strand specific libraries.
I don't know how the two compare at small input quantities.
And I don't know the best reagents for the miRNA approach. Would probably buy a kit. Would people just use Illumina's?
Any feedback would be very much appreciated. Thanks!
We can do this by randomly-primed RT, second strand synth, end prep and adaptor ligation, a la standard RNASeq.
Or we can do it by sequential ssRNA adaptor ligation, a la miRNASeq.
Which would people advise using?
In favor of the standard approach is that we do this method all the time.
In favor of the miRNA approach is that it gives strand specific libraries.
I don't know how the two compare at small input quantities.
And I don't know the best reagents for the miRNA approach. Would probably buy a kit. Would people just use Illumina's?
Any feedback would be very much appreciated. Thanks!
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