TRIzol Reagent

RNA isolation / purification Cells - immortalized RPMI 2650

Experiment
RNA isolation / purification Cells - immortalized RPMI 2650
Product
TRIzol Reagent from Thermo Fisher Scientific
Manufacturer
Thermo Fisher Scientific

Protocol tips

Protocol tips
- For low 260/230 readings the best approach is to try washing sample (precipitation) with ethanol to desalt it.

Publication protocol

Total RNA was isolated using the Trizol reagent (Invitrogen, USA) according to the manufacturer’s instructions. Total RNA was incubated with DNase I for 15 min at 37°C. One microgram of total RNA was applied into reverse transcription reaction using First Strand cDNA Synthesis Kit (Roche). One microliter of cDNA was amplified in a total reaction volume of 20 μl containing 100 nM gene specific primer and 10 μl of 2× SYBR® Green PCR Master Mix (Applied Biosystems) using StepOne Real-Time System (Applied Biosystems). The Q-PCR experiment was performed in triplicate and the mean values were used for calculations. The IGFBP-7 mRNA expression index was normalized with the reference gene GADPH using the comparative Ct method (2-△CT)

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Discussion

Discussion

5 years ago

Author: Switzerland

I am having trouble using the QiageN RNAeasy mini kit for CD34+ cells. Any tips?

I am having trouble using the QiageN RNAeasy mini kit for CD34+ cells. Any tips?

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Papers

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for TRIzol Reagent below.

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