TRI Reagent™ Solution

Protocol tips

Upstream tips	Protocol tips	Downstream 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.

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).

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

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.

Manufacturer protocol

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

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing RNA isolation / purification Cells - immortalized K-562 using TRI Reagent™ Solution from Thermo Fisher Scientific. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.