Small RNA seq of Stolons-7 days post LD induction
Small RNA libraries for sequencing were constructed at the Genotypic Technology s Genomics facility according to the Illumina TruSeq Small RNA library protocol outlined in  TruSeq Small RNA Sample Preparation Guide  (Part # 15004197; Rev. G; January 2015). 1 g of Total RNA was used as the starting material. Briefly, 3  adaptors were ligated to the specific 3 OH 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 libraries were size selected in the range of 140 bp  160 bp followed by overnight gel elution and salt precipitation using Glycogen, 6M Sodium Acetate and Absolute Ethanol. The precipitate was re-suspended in Nuclease free water. The prepared libraries were quantified using Qubit Fluorometer, and validated for quality by running an aliquot on High Sensitivity Bioanalyzer Chip (Agilent). The adapter ligated molecules were quantified on qPCR using KAPA library quantification kit (KAPA Biosystems).