GSM1704907: PbWT DR mouse1; Plasmodium berghei; RNA-Seq
The total mRNA was extracted using the Trizol reagent and Magnetic mRNA Isolation Kit (NEB) according to manufacturer instructions First strand cDNA was synthesized with Random Hexamer primers and SuperScript III Reverse Transcriptase (Life Technologies), following the manufacturer’s instructions. The DNA/RNA hybrids were purified using Agencourt AMPure XP beads and the 2nd strand was synthesized using dUTP nucleotide mix, DNA Polymerase I (E. coli) and RNAse H (NEB) by 2.5 hours of incubation at 16°C. CDNA was fragmented to approximately 200 nucleotides using Covaris S220 System. The fragments were end-repaired, TA-tailed and ligated to Illumina adaptors with index barcodes using NEBNext kits according to manufacturer instructions. Excess adaptors were removed with 2 rounds of clean up with 50μl of Agencourt AMPure XP beads (Beckman Coulter). Final libraries were eluted in 30μl water, visualized on an Agilent Bioanalyzer 2100 High Sensitivity DNA chip and quantified by qPCR. A pool of the indexed libraries was sequenced on an Illumina HiSeq2000, with 100bp paired-end reads.
GEO Accession
GSM1704907