Caragana korshinskii sequencing project- nt_ck
Examination of specific stress-resistance-related genes in drought-treated or salt-treated seedling plants of C. korshinskii.Total RNA was extracted from C. korshinskii whole seedlings treated with 400 mM NaCl or drought stress using a Plant RNA Extraction Kit (Autolab, China), following the manufacturer’s protocol. The concentration and quality of each RNA sample were determined using a NanoDrop 2000TM micro-volume spectrophotometer (Thermo Scientific, Waltham, MA, USA) and gel electrophoresis. The poly (A) mRNA was isolated from 10 mg total RNA using magnetic oligo(dT) beads, then divided into short fragments by fragmentation buffer (Ambion, Austin, TX, USA). The first-strand cDNA was synthesized by random-hexamer primers, and then the second-strand cDNA was synthesized using DNA polymerase I (New England BioLabs), RNase H (Invitrogen), buffer, and dNTPs. Short DNA fragments were purified with a QIAquick PCR Purification Kit (Qiagen Inc., Valencia, CA, USA), then subjected to end repair and poly(A) addition and ligated to sequencing adaptors. Suitable fragments were purified by agarose-gel electrophoresis and gathered by PCR amplification. Three cDNA libraries were then constructed and sequenced using the Illumina HiSeq™ 2000 platform.