ProtoScript® II First Strand cDNA Synthesis Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Treat sample with DNase I in the presence of 20 units of RNaseOUT at 37°C for 20 min. Inactivate DNAse with 25 mM EDTA at 80°C for 2 min

Upstream tips
Treat sample with DNase I in the presence of 20 units of RNaseOUT at 37°C for 20 min. Inactivate DNAse with 25 mM EDTA at 80°C for 2 min

Publication protocol

Total RNA was extracted by hot acidic phenol method as described [68]. 20 μg of RNA was treated with 10 units of DNase I (Clontech) in the presence of 20 units of RNaseOUT (Invitrogen) at 37°C for 20 min. DNase I was inactivated by incubation with 25 mM EDTA at 80°C for 2 min. cDNA was synthesized using ProtoScript II First Strand cDNA Synthesis Kit (New England BioLab) according to the manufacturer’s instruction. Real-time PCR was performed as previously described [55]. PCR amplification from MEC1 and TEL1 transcript was normalized using that from ACT1 transcript. PCR primers are listed in S2 Table.

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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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