GSM1635583: LV-GAS5-mRNA; Homo sapiens; RNA-Seq
Total RNAs were harvested using Trizol reagent and small RNAs were harvested using miRcute miRNA extraction reagent (Tian Gen, China). RNA-seq: The complementary DNA (cDNA) libraries for single-end sequencing were prepared using Ion Total RNA-Seq Kit v2.0 (Life Technologies) according to the manufacturer’s instructions. The cDNA libraries were then processed for the Proton Sequencing process according to the commercially available protocols. Samples were diluted and mixed, the mixture was processed on a OneTouch 2 instrument (Life Technologies) and enriched on a OneTouch 2 ES station (Life Technologies) for preparing the template-positive Ion PI™ Ion Sphere™ Particles (Life Technologies) according to Ion PI™ Template OT2 200 Kit v2.0 (Life Technologies). After enrichment, the mixed template-positive Ion PI™ Ion Sphere™ Particles of samples was loaded on to 1 P1v2 Proton Chip (Life Technologies) and sequenced on Proton Sequencers according to Ion PI Sequencing 200 Kit v2.0 (Life Technologies). miRNA-seq: The complementary DNA (cDNA) libraries for single-end sequencing were prepared using TruseqTM Small RNA sample prep Kit (Illumina) according to the manufacturer’s instructions. 1ug small RNA was used to add 3’ adaptor and 5’ adaptor to small RNA and RT-PCR was applied to the small RNA with adaptor to achieve the 1st cDNA; Truseq (Illumina) primer was used for PCR enrichment and introducing index sequence. Size selection was applied to obtain the 145-160bp library sequence. TBS380(Picogreen) was introduced to quantify the library concentration. We applied the cluster formation on flow cell utilizing cBot and sequencing on Hiseq2000 Platform.
GEO Accession
GSM1635583