small RNA sequencing
1 g of RNA was 3 adaptors were ligated to the specific 3OH group of micro RNAs followed by 5 adaptor ligation. The ligated products were reverse transcribed with Superscript III Reverse transcriptase by priming with Reverse transcriptase primers. The cDNA was enriched and barcoded by PCR (15 cycles) and cleaned using Polyacrylamide gel. The library was size selected in the range of 140  160 bp followed by overnight gel elution and salt precipitation using Glycogen, 3M Sodium Acetate and Absolute Ethanol. The precipitate was re-suspended in Nuclease free water. The prepared library was quantified using Qubit Fluorometer, and validated for quality by running an aliquot on High Sensitivity Bioanalyzer Chip.
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