title: GSM1289020: HepG2- PBANKA-WT-mRNA Sequencing; Homo sapiens; RNA-Seq
library_construction_protocol: RNA was isolated from FACS sorted parasitized HepG2 cells using Trizol method and concentration of RNA determined by Qubit flourimeter (Invitrogen) using QubitTM RNA Assay Kit. The total RNA for each sample was analyzed on Agilent bioanalyzer using Agilent RNA 6000 Pico kit. Libraries were prepared according to Illumina's instructions accompanying theTruseq standed RNA sample preparation kit (FC-122-1001). Briefly, the poly-A containing mRNA molecules were purified from total RNA using poly-T oligo-attached magnetic beads. The mRNA was fragmented into small pieces, using divalent cations under elevated temperature, and converted into first strand cDNA using reverse transcriptase and random primers. The second strand of cDNA was synthesized using DNA polymerase-I and RNAse-H enzymes. The synthesized cDNA have an additional ‘A’ base, which is required for adapter ligation. The products were purified and enriched with PCR to create the final cDNA library. The cDNA library for each sample was analyzed on Agilent 2100 Bioanalyzer using Agilent DNA 1000 kit.
attributes.tag: GEO Accession
attributes.value: GSM1289020