GSM1240331: cDNA from RNAs of Bupleurum scorzonerifolium roots; Bupleurum scorzonerifolium; RNA-Seq
Total RNA was isolated using Trizol reagent® according to instructions of manufacturer (Invitrogen, Karlsruhe, Germany) and then mRNA was purified and enriched using the mRNA purification kit (Promega, Fitchburg, USA). The mRNAs were reverse-transcribed using Powerscript™ II (TaKaRa, Japan) with PCR primers SMART IV™ Oligonucleotide (5'-AAGCAGTGGTATCAACGCAGAGTGGCCATTACGGCCGGG-3') and CDS III/3' PCR Primer (5'-ATTCTAGAGGCCGAGGCGGCCGACATG-d(T)30N-1N-3'). Long Distance PCR for double strand cDNA amplification was performed with LA Taq enzyme (TaKaRa, Japan) for 25 cycles (95°C for 30 s, 68°C for 8 min) according to the SMART™ cDNA Library Construction Kit User Manual (Clontech, CA, USA). Finally, double strand cDNAs were purified using DNA purification kit (Qiagen, Venlo, The Netherlands) to generate high quality cDNA. Approximately 10 μg of sheared cDNA were used for GS20-454 sequencing. A total sequencing runs were performed with B. chinense a half and B. scorzonerifolium a half.
GEO Accession
GSM1240331