title: GSM2982076: T2_miRNA_1; Arachis hypogaea; miRNA-Seq
library_construction_protocol: Fresh seeds were harvested from plants of both peanut RILs at 15 DAF and 35 DAF, and RNA was extracted using TRK-1001,TruSeq Small  RNA Sample Prep Kits was used with 1 ug of total RNA for the construction of sequencing libraries. Total RNA was ligated to the RNA 3' and RNA 5' adapters and reverse transcription, followed by PCR, was performed to make cDNA constructs of the small RNAs. The small cDNA fractions that ranged from 22 nt to 30 nt in length were then isolated via 6% denaturing polyacrylamide gel electrophoresis. Finally, the cDNA constructs were purified, and the library was validated. We then performed single-end sequencing (50 bp) on an Illumina Hiseq2500 following the vendor's recommended protocol.  Approximately 20 ug of total RNA were used to prepare Degradome library. (1) Approximately 150 ng of poly(A)+ RNA was used as input RNA and annealing with Biotinylated Random Primers. (2) Strapavidin capture of RNA fragments through Biotinylated Random Primers. (3)5' adaptor ligation to to only those RNAs containing 5'-monophosphates. (4) Reverse transcription and PCR (5) Libraries were sequenced using the 5'adapter only, resulting in the sequencing of the first 47 nucleotides of the inserts that represented the 5' ends of the original RNAs.
attributes.tag: GEO Accession
attributes.value: GSM2982076