ProtoScript® II 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 Mixand 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 Mixand M-MuLV Enzyme Mix and incubate reaction at 42°C for one hour. Inactivate the enzyme at 80°C for 5 minutes

Publication protocol

LacZ staining and whole-mount in situ hybridization was carried out following standard protocols. For RT-qPCR, total RNA was extracted from the frozen tissues using RNeasy kit (QIAGEN), and then cDNA was synthesized using the ProtoScript II First Strand cDNA Synthesis Kit (New England Biolabs). The quantitative PCR was performed using StepOne Real-Time PCR System with SYBR green reagent (Applied Biosystems). Gapdh was used to normalize expression level for each sample. The extra-embryonic membranes were used for PCR-genotyping of the embryos.

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Manufacturer protocol

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

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