NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina®

Protocol tips

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	Starting Material: 100 ng-1μl total RNA in a 12 μl total volume.

Protocol tips
Starting Material: 100 ng-1μl total RNA in a 12 μl total volume.

Publication protocol

Total RNA was isolated as described above from control, sh73/DLK- and sh69/DLK-depleted cells. The quality of total RNA was evaluated using the Agilent 2100 BioAnalyzer and the Agilent RNA 6000 Nano Kit (#5067-1511) according to the manufacturer’s instructions. For all samples, RNA integrity numbers were sufficiently high (>9.5) to perform mRNA sequencing. mRNA was purified from 5 μg of total RNA using the NEBNext Poly(A) mRNA Magnetic Isolation Module (#E7490, New England Biolabs, Whitby, Ontario) following manufacturer’s instructions. cDNA libraries were then prepared with 25 ng mRNA of each sample using the NEBNext Ultra Directional RNA Library Prep Kit for Illumina (#E7420, New England Biolabs) and barcoded by PCR for subsequent multiplexed sequencing. The quality and quantity of the librairies were assessed using the Agilent High Sensitivity DNA Kit (#5067-4626) on Agilent 2100 BioAnalyzer. Sequencing of the multiplexed cDNA libraries was performed on an Illumina HiSeq 2500 system (Illumina, San Diego, CA, USA) in two lanes at the Institut de Recherches Cliniques de Montréal (IRCM).

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Manufacturer protocol

Download the product protocol from New England BioLabs for NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina® below.

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