GSM1709502: Degradome long-time; Gossypium hirsutum; RNA-Seq
Total RNA was isolated using a modified CTAB method and purified using Qiagen RNeasy columns (Qiagen, Hilden, Germany).By using the Oligotex kit (Qiagen, location), 200 μg of total RNA was used for extracting poly (A) RNA, which were ligated with an RNA adapter consisting of a MmeI recognition site in its 3′end. The extracted PloyA RNAs  were ligated with an RNA adapter consisting of a MmeI recognition site in its 3′end. After ligation, first-strand cDNA was generated using oligod (T) and the PCR product was amplified using five PCR cycles. The PCR product was purified and digested with MmeI. The digested PCR product was then ligated to a double-stranded DNA oligonucleotide with degenerate nucleotides at the 5′-or 3′-ends. The ligation product was further gel purified and amplified using 10 PCR cycles. The final PCR product was purified and sequenced using Illumina’s sequencing by synthesis (SBS) sequencing technology.