GSM8265152: 1_M90T_30C_S8; Shigella flexneri; RNA-Seq
Total RNA was extracted with acidic phenol, DnaseI treated and ribosomal RNA were depleted. An oligonucleotide adapter was ligated to the 3' end of the RNA molecules. First-strand cDNA synthesis was performed using M-MLV reverse transcriptase and the 3' adapter as primer. 5' Illumina TruSeq sequencing adapter was ligated to the 3' end of the antisense cDNA. The resulting cDNA was PCR-amplified to about 10-20ng/μl using a high-fidelity DNA polymerase. The cDNA was purified using the Agencourt AMPure XP kit (Beckman Coulter Genomics).