RNA-Seq of alfafa shoot under phosphate deficiency stress
Total RNA was extracted from approximately 100 mg ground powder based on the method of TRIzol manufacturers instruction. Library construction was processed using Illumina TruseqTM RNA sample prep Kit. Simply, The mRNA containing the 3 '-terminal ployA tail structure was aggregated using magnetic beads and randomly fragmented into 300bp fragments. Then mRNA fragments were transcribed into the first strand cDNA. Following the double-strand cDNA were synthesized and purified. Adaptors were ligated after adenylation the 3 ends. Finally, the fragments were purified by 2% agarose gel electrophoresis and enriched by PCR amplification. cDNA libraries were validated. Products were sequenced across using 2  150 bp chemistry by HISEQ.