GSM1541985: TM-wt-glucose-1; Thermotoga maritima MSB8; RNA-Seq
Total RNA was isolated via enzymatic lysis followed by purification with Qiagen's Rneasy Kit with on column Dnase treatment. Strand-specific paired-end librariries were constructed using the "dUTP" methods. Breifely, rRNA depletion was performed using Epicentre's RiboZero kit. Depleted RNA was then fragmented, primed with random hexamers, and reverse transcribed. Second strand synthesis was performed with incorporation of dUTPs instead of dTTPs. This creates a marker on the second strand for enzymatic degradation of this strand producing strand specific libraries. After second strand synthesis, end-repair, dA tailing, adaptor ligation, USER treatment and PCR enrichment were performed.