TRI Reagent® MRC

RNA isolation / purification Tissue - Human Placenta

Experiment
RNA isolation / purification Tissue - Human Placenta
Product
TRI Reagent® MRC from Molecular Research Center, Inc.
Manufacturer
Molecular Research Center, Inc.

Protocol tips

Upstream tips
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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Discussion

Discussion

4 years ago

Author: Paul G. Macon United States

RNA isolation from tissue

How do I extract RNA from animal tissue without using liquid nitrogen? I tried the RNA extraction by using the TRIzol reagent and I homogenize the tissue using polytron homogenizer at room temperature for 30secs is this correct?

Discussion

5 years ago

Author: Aaron Stege Netherlands

Problem in phase separation after using serum/plasma kit

I used a serum/plasma kit for my serum samples. After the phase separation the samples should have 3 phases: a colourless aqueous phase, a white interphase and a red organic phase. However, in some of my samples there was no aqueous phase unless I wait for an extended period of time. How can I circumvent this problem?

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Papers

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Manufacturer protocol

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

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