PrimeScript™ 1st strand cDNA Synthesis Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	After DNAse treatment add Oligo dT Primer,dNTP Mixture,Template RNA and RNase-free dH O and heat at 65°C for 5 min. Add reaction mix to the above mix and incubate at 30°C 10 min* and 42 to 50°C for 30 to 60 min

Protocol tips
After DNAse treatment add Oligo dT Primer,dNTP Mixture,Template RNA and RNase-free dH O and heat at 65°C for 5 min. Add reaction mix to the above mix and incubate at 30°C 10 min* and 42 to 50°C for 30 to 60 min

Publication protocol

All M. tuberculosis strains were sub-cultured in 7H9 medium, supplemented with Albumin Dextrose Catalase (ADC), in the absence of rifampin (control strains) and presence of sub-MIC of rifampin (MIC = 128 mg/L), and collected after four weeks for RNA extraction. Total bacterial RNA was isolated, using PREP-NA DNA/RNA extraction kit (DNA technology, Russia). The quality and integrity of the total RNA was assessed, using a nanophotometer (Thermoscientific, USA) and agarose gel electrophoresis. After DNase I treatment, RNA (1.5 μg) was reverse transcribed, according to the manufacturer’s recommendations (PrimeScript First Strand cDNA Synthesis Kit, Takara, Japan), and the thermal cycling conditions were as follows: 25 °C for 10 min, 42 °C for 60 min, and 85 °C for 5 min. The cDNAs were maintained at −20 °C, until further use

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Manufacturer protocol

Download the product protocol from Takara Bio Inc for PrimeScript™ 1st strand cDNA Synthesis Kit below.

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