S5_E7_0_neg
500 nanograms of RNA were used as input to the Illumina TruSeq Stranded mRNA Library Prep Kit (Illumina, 20020595). The libraries were generated per manufacturers specifications, except Illumina adapters provided with the kit were replaced with NEXTFLEX Unique Dual Index Barcodes (Perkin Elmer, references NOVA-514150, NOVA-514151, NOVA-514152, NOVA-514152). RNA-Seq libraries were amplified using the Illumina Truseq primer mix. The PCR amplified RNA-Seq library products were then quantified using the Quant-iT dsDNA Assay Kit, High Sensitivity (In Vitrogen, Q33120) on a Gemini XPS (Molecular Devices, XPS05542). The library profiles were checked using High Sensivity NGS Fragment Analysis kit (1-6000bp) (Agilent Technologies, DNF-474-1000) on the 5300 Fragment Analyzer System. The libraries were diluted to 5 nanomolar in Elution Buffer, pooled, denatured, and loaded at 300 picomolar on Illumina NovaSeq 6000 flowcells SP or S1, using NovaSeq 6000 Reagent Kit v1.5. The RNA-Seq libraries were sequenced 50 or 100 base pair paired-end with 8 base dual indexes. The sequence intensity files were generated on instrument using the Illumina Real Time Analysis software. Paired-end 50 or 100 base pair reads were generated for an average of about 50 million or 90 million read pairs per sample. FASTQ files were extracted using Illuminas bcl2fastq software (v2.17.1.14)
SUB15380681
GX_IL_LIB_22_A509