ProtoScript® First Strand cDNA Synthesis Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Mix RNA sample and primer d(T)23VN and denature RNA for 5 minutes at 70°C. Add M-MuLV Reaction Mix and M-MuLV Enzyme Mix and incubate reaction at 42°C for one hour. Inactivate the enzyme at 80°C for 5 minutes

Protocol tips
Mix RNA sample and primer d(T)23VN and denature RNA for 5 minutes at 70°C. Add M-MuLV Reaction Mix and M-MuLV Enzyme Mix and incubate reaction at 42°C for one hour. Inactivate the enzyme at 80°C for 5 minutes

Publication protocol

Total RNA was extracted from 5 ml of cells grown to mid-exponential phase using the Ambion RiboPure-Bacteria kit and treated with DNase I for 2 h at 37 °C. One microgram of this RNA was reverse transcribed with random primers using the Protoscript First Strand cDNA Synthesis kit (New England Biolabs). Negative controls were also performed without reverse transcriptase to ensure a lack of genomic DNA contamination. The cDNA was then used in a PCR with gene-specific primers designed to amplify approximately 300 bp regions.

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Manufacturer protocol

Download the product protocol from New England BioLabs for ProtoScript® First Strand cDNA Synthesis Kit below.

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