TRI Reagent™ Solution

RNA isolation / purification Cells - immortalized K-562

Experiment
RNA isolation / purification Cells - immortalized K-562
Product
TRI Reagent™ Solution from Thermo Fisher Scientific
Manufacturer
Thermo Fisher Scientific

Protocol tips

Upstream tips
- 1-Bromo-3-chloropropane is less toxic than chloroform and its use for phase separation decreases the possibility of contaminating RNA with DNA.

- The chloroform used for phase separation should not contain isoamyl alcohol or other additives.

Publication protocol

Total RNA isolation was performed by cell lysis with TRI-Reagent (Ambion, Foster City, USA) by taking the upper aqueous phase obtained after centrifugation at 1000g for 10 min. RNA was then washed in isopropanol and cool 75% ethanol, resuspended in nuclease-free water and kept at -20°C for further analysis.

MicroRNA isolation was performed by miRCURY™ RNA Isolation Kit—cell & plant (EXIQON, Vedbaek, Denmark) according to manufacturer’s instruction.

cDNA from total RNA extracted in TRI-Reagent was then synthesized using the iScript™ cDNA Sinthesis Kit (Bio-Rad Laboratories, Hercules, USA) and qRT-PCR analysis of Bax, Bcl-2, BNIP3, SMO and Gli1 RNA expression was performed on cDNAs by using iQ™ SYBR Green Supermix (Bio-Rad Laboratories, Hercules, USA).

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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

Check out relevant papers found by Labettor's AI that are relevant for performing RNA isolation / purification Cells - immortalized K-562 using TRI Reagent™ Solution from Thermo Fisher Scientific.

Paper title
Novel Acylguanidine Derivatives Targeting Smoothened Induce Antiproliferative and Pro-Apoptotic Effects in Chronic Myeloid Leukemia Cells
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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for TRI Reagent™ Solution below.

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