KAPA RNA HyperPrep Kit with RiboErase (HMR)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Input RNA: 1 ng-100 ng.

Protocol tips
Input RNA: 1 ng-100 ng.

Publication protocol

For paRNA-seq, we purified poly(A) RNA from 1 µg of ES total RNA using a NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB). For rdRNA-seq, we depleted rRNA from 1 µg of ES total RNA using a GeneRead rRNA Depletion Kit (Qiagen). We prepared conventional RNA-seq library DNA with the resulting RNA using a commercial kit (NEBNext Ultra Directional RNA Library Prep Kit for Illumina; NEB) in accordance with the manufacturer’s protocol, with slight modifications during RT and PCR steps. We used SuperScript III (Thermo Fisher) for RT. We used KAPA HiFi DNA polymerase (Kapa Biosystems) in the PCR step. When we performed rdRNA-seq using 10 or 1 ng of ES total RNA, we utilized a KAPA RNA HyperPrep Kit with RiboErase (Kapa Biosystems) for the preparation of sequencing library DNA according to the manual instructions.

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Manufacturer protocol

Download the product protocol from Roche Lifesciences for KAPA RNA HyperPrep Kit with RiboErase (HMR) below.

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