TRI Reagent® MRC

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This kit purifies RNA from up to 50-100 mg tissue

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This kit purifies RNA from up to 50-100 mg tissue

Publication protocol

Total RNA was isolated from 50 to 100 mg of tissue by homogenising in cold Tri Reagent® (Helena Biosciences, UK) on wet ice using a Dounce homogeniser, precipitating in isopropanol and dissolving in DEPC treated water. RNA quantity and purity were measured spectrophotometrically by measuring OD 260/280 and OD 260/230 in a NanoDrop™ 2000c spectrophotometer (Thermo Scientific). RNA integrity was determined using Agilent 2100 Bioanalyzer® (Agilent Technologies) according to the manufacturer's instructions.

1 μg RNA was Dnase treated with Dnase I, Amp Grade® (Invitrogen) according to the manufacturer's instruction. Real time PCR reactions for transferrin receptor 1 (TfR1) (primer catalogue number QT00094850) were run using the 7500 Fast Real Time PCR system (Applied Biosystem) with ubiquitin (QT00234430) as the housekeeping gene. Gene expression was normalised to ubiquitin and expressed as 2−ΔCT. Single melt curves were obtained for all samples, indicating that a single PCR product was generated.

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Manufacturer protocol

Download the product protocol from Molecular Research Center, Inc. for TRI Reagent® MRC below.

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