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This protocol describes the TruSeq stranded mRNA preparation and sequencing for RNA-Seq experiments. The protocol is primarily focused on the generation of a stranded mRNA-Seq library from total RNA or polyA RNA extracts. While the steps are similar, data generated from both RNA fractions produce unique quality control metrics that need to be considered. This protocol is based on the two-step protocol developed by OATG (Illumina) for the preparation of TruSeq stranded mRNA libraries from total RNA extracts. The protocol also contains some variations, such as the duplex repair and the Enrichment of 3′ poly(A) fraction, which can be adapted to adapt the protocol to other mRNA-Seq kits.Purification and characterization of recombinant human lymphotoxin-α and its use in a bioassay for tumors of epithelial origin.
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