HCV patients with Cirrhosis
Small RNA libraries for sequencing were constructed at the Genotypic TechnologysGenomics facility according to the Illumina TruSeq Small RNA library protocol outlined inTruSeq Small RNA Sample Preparation Guide (Part # 15004197; Rev. F; February 2014).75 ng of Total RNA was used as the starting material. Briefly, 3 adaptors were ligated to thespecific 3OH group of micro RNAs followed by 5 adaptor ligation. The ligated products werereverse transcribed with Superscript III Reverse transcriptase by priming with Reversetranscriptase primers. The cDNA was enriched and barcoded by PCR (21 cycles) andcleaned using Polyacrylamide gel. The library was size selected in the range of 140  160bpfollowed by overnight gel elution and salt precipitation using Glycogen, 3M Sodium Acetateand Absolute Ethanol. The precipitate was re-suspended in Nuclease free water. Theprepared library was quantified using QubitFluorometer, and validated for quality by runningan aliquot on High Sensitivity Bioanalyzer Chip (Agilent).Adapter sequences:Adapter 1: AATGATACGGCGACCACCGAGATCTACACGTTCAGAGTTCTACAGTCCGAAdapter 2: CAAGCAGAAGACGGCATACGAGAT[index]GTGACTGGAGTTCCTTGGCACCCGAGAATTCCA
SUB9280008
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