Nebnext multiplex oligos for index primer 1 12

Messenger RNA (mRNA) was enriched with a NEBNext Poly (A) mRNA Magnetic Isolation Module (NEB, E7490, Ipswich, MA). Using the enriched mRNA as template, cDNA libraries were constructed using the NEBNext Ultra RNA Library Prep Kit Illumina (NEB, E7530, Ipswich, MA) and NEBNext Multiplex Oligos for Illumina (Index Primer 1–12) (NEB, E7600, Ipswich, MA) following the manufacturer’s instructions. To verify the quality, DNA concentration and product size of the cDNA libraries, a Qubit 2.0 Fluorometer (Life Technologies, Q32866), Qubit dsDNA BR assay kit (Life Technologies, Q32850), High Sensitivity DNA Analysis Kit (Agilent, 5067–4626) and Bioanalyzer were used. cDNA libraries were sequenced on an Illumina MiSeq Platform employing a 150 base pair single-end NGS setting. Data from MiSeq sequencing runs were uploaded and stored in BaseSpace (https://basespace.illumina.com) for data analysis.

Messenger RNA was isolated with a NEBNext Poly (A) mRNA Magnetic Isolation Module (New England Biolabs, E7490). The enriched mRNA served as template for cDNA library preparation with a NEBNext Ultra RNA Library Prep Kit Illumina (NEB, E7530) and NEBNext Multiplex Oligos for Illumina (Index Primer 1–12) (NEB, E7600) following the manufacturer’s instructions. Assessment of quality, concentration and size of the cDNA, was performed for each library using a Qubit 2.0 Fluorometer (Life Technologies, Q32866), Qubit dsDNA BR assay kit (Life Technologies, Q32850), High Sensitivity DNA Analysis Kit (Agilent, 5067–4626) and Agilent 2100 Bioanalyzer. cDNA libraries were sequenced on an Illumina MiSeq Platform employing a 150 base pair paired-end NGS setting. Sequencing data were uploaded and stored in BaseSpace (https://basespace.illumina.com) for subsequent analysis.