title: GSM2267310: ∆sRNA154 replicate 2; Methanosarcina mazei Go1; RNA-Seq
library_construction_protocol: RNA isolation was performed using TRI reagent following the manufacturer´s protocol followed by DNaseI treatment and phenol-chloroform precipitation For cDNA synthesis both RNA fractions were poly(A)-tailed using poly(A) polymerase. This was followed by treatment with TAP to remove the 5' PPP structures. Afterwards, an RNA adapter was ligated to the 5'-monophosphate of the RNAs. First strand cDNA synthesis was performed using an oligo(dT)-adapter primer and the M-MLV reverse transcriptase. The resulting cDNAs were PCR-amplified to about 10-20 ng/μl using a high fidelity DNA polymerase. PCR cycles performed and barcode sequences, which are part of the 3' sequencing adapter. The cDNAs were purified using the Agencourt AMPure XP kit (Beckman Coulter Genomics) and were analyzed by capillary electrophoresis. For Illumina sequencing, the cDNAs were pooled in approximately equimolar amounts.
attributes.tag: GEO Accession
attributes.value: GSM2267310