SuperScript® II Reverse Transcriptase

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Heat Oligo mixture to 65°C for 5 min and quick chill on ice and later add reaction components. Mix and incubate at 42°C for 2 min. Add 1 μL (200 units) of SuperScript™ II RT and mix by pipetting gently up and down. Incubate at 42°C for 50 min. Inactivate the reaction by heating at 70°C for 15 min.

Protocol tips
Heat Oligo mixture to 65°C for 5 min and quick chill on ice and later add reaction components. Mix and incubate at 42°C for 2 min. Add 1 μL (200 units) of SuperScript™ II RT and mix by pipetting gently up and down. Incubate at 42°C for 50 min. Inactivate the reaction by heating at 70°C for 15 min.

Publication protocol

Total RNA were extracted at the designated time points in cell cycle with SV Total RNA Isolation System (Promega). For qPCR, 1 mg RNA was converted into cDNA with Superscript II Reverse Transcriptase (Invitrogen), and Q-PCR was performed with Go-Taq qPCR Master Mix (Promega) on a Step One Plus device (Applied biosystems) for 40 cycles (95C, 15 sec; 55C, 30 sec; 72C, 30 sec). Primer sequences are listed in Table S2.

For deep-sequencing, RNA concentration was measured with Nanodrop and integrity was assessed on Bioanalyzer 2100 (Agilent) using RNA 6000 pico kit (Agilent), according to the manufacturer’s protocol. For each time point, RNA from 2 independent experiments were pulled into 1 sample and deep sequenced on SOLID sequencing platform at Uppsala Genome Center.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for SuperScript® II Reverse Transcriptase below.

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