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Old 09-01-2014, 03:03 AM   #2
nucacidhunter
Jafar Jabbari
 
Location: Melbourne

Join Date: Jan 2013
Posts: 1,177
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Small RNA library preparation involves ligating adapters to termini of RNA and PCR after RT reaction. The reaction condition is optimised for amplification of small RNA species, so the larger RNA fractions will not amplify efficiently. In addition, there is a gel size selection step which cuts fragments generated from small RNA fraction for sequencing. RNAseq libraries are prepared by random priming of fragmented RNA, so the small RNA will not have much chance of primer binding and reverse transcription. If the aim is to study both small RNA and total RNA-rRNA or mRNA from the same sample, separate libraries have to be prepared for each.
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