GSM3167439: sp 1; Synechocystis sp. PCC 6803; RNA-Seq
Cells were disrupted using the FastRNA Pro Blue Kit (MP Bio). Briefly, cell pellets were resuspended in 1 ml of the supplied phenol lysis solution and added to the Lysing Matrix before disruption in a Precellys 24 homogenizer (Bertin Technologies). After centrifugation, the supernatant was removed and chloroform extracted. Total RNA was then isolated from the aqueous phase using the Total RNA Purification Plus Kit (Norgen). Ribosomal RNA molecules were depleted using the Ribo-Zero rRNA Removal Kit (Illumina). Samples were fragmented using ultrasound, adapter was ligated to the 3' end of the RNA. First-strand cDNAsynthesis was performed using PrimeScript Reverse Transcriptase (TaKaRa). cDNA was purified and the 5' Illumina TruSeq sequencing adapter was ligated. cDNA was PCR-amplified to about 10-20 ng/μl using a high fidelity DNA polymerase. The cDNA was purified using the Agencourt AMPure XP kit (Beckman Coulter Genomics).
GEO Accession
GSM3167439