KAPA Stranded mRNA-Seq Kit

Protocol tips

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	This protocol has been validated for library construction from 100 ng – 4 μg intact, total RNA, in ≤50 µL of RNase-free water

Protocol tips
This protocol has been validated for library construction from 100 ng – 4 μg intact, total RNA, in ≤50 µL of RNase-free water

Publication protocol

Total RNA was isolated from PBMCs using RNeasy (Qiagen) or Arcturus PicoPure (Life Technologies) kits following the manufacturer’s protocols. During RNA isolation, DNase treatment was additionally performed using the RNase-free DNase set (Qiagen). RNA quality was checked using an Agilent 2100 Expert bioanalyzer (Agilent Technologies). RNA quality was reported as a score from 1 to 10, and samples falling below the threshold of 8.0 were omitted from the study. cDNA libraries were prepared using a TruSeq Stranded Total RNA LT Sample Prep kit with Ribo-Zero Gold (Illumina), a Kapa Stranded mRNA-Seq Library Prep kit (Kapa Biosystems), or NuGEN Ovation RNA-seq v2 (NuGEN) according to the manufacturer’s instructions using 100 or 500 ng of total RNA. Final libraries were analyzed on a Bioanalyzer DNA 1000 chip (Agilent Technologies). Paired-end sequencing (2 × 75 bp or 2 × 100 bp) of stranded total RNA libraries was performed in an Illumina HiSeq2500 using SBS v3 sequencing reagents.

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Manufacturer protocol

Download the product protocol from Roche Lifesciences for KAPA Stranded mRNA-Seq Kit below.

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