GSM4850495: Teosinte_DroughtSubmergence; Zea mays subsp. parviglumis; miRNA-Seq
The shoots were removed, frozen in liquid nitrogen and processed with TRIzol to extract total RNA. The small RNA fraction of 20-30 nt was size-purified from total RNA on denaturing polyacrylamide gels. Small RNA fractions purified from 12µg of total RNA were ligated to a preadenylated adapter in the 3'end. Next,  the 5' RNA Illumina adapter was ligated to the 5'end of small RNAs. The products with adapters in both ends were used in cDNA synthesis reactions followed by a 15 cycle PCR with primers compatible with Illumina. The products were purified, quantified, and sequenced on the Illumina HiSEQ2500.