RNeasy Plus Mini Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
		- Include DNAse treatment for 15-20min. - Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C. - Use water to elute the RNA that is warmed to ~60`C.

Downstream tips
- Include DNAse treatment for 15-20min. - Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C. - Use water to elute the RNA that is warmed to ~60`C.

Publication protocol

The total RNA content, including miRNAs, was isolated from the transduced and control cells using the RNX plus reagent (CinnaGen, Tehran, Iran) according to the manufacturer’s instructions, 48 hours after the transduction. The RNA extracts were kept at −80°C until use. Next, 5 μg of total RNA, used as a template, was polyadenylated with poly(A) polymerase enzyme. Complementary DNA (cDNA) was synthesized using a cDNA synthesis kit (Fermentas, Massachusetts, USA) and specific primers. The sequence-specific RT-qPCR primers for miR-192 and endogenous control SNORD were purchased from Bonyakhteh Research Center, Iran. RT-qPCR analysis was carried out on the Rotor-gene 6000 real-time PCR device (Corbett, Mortlake, Australia) using Taq DNA Polymerase Master Mix (Ampliqon, Rodovre, Denmark), and the following PCR conditions were applied: 95°C for 10 minutes and then 95°C for 15 seconds, 60°C for 60 seconds for up to 40 cycles (n = 3). The gene expression cycle threshold (∆∆Ct) values of miRNAs were calculated after normalizing with SNORD internal control.

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 NALM-6 using RNeasy Plus Mini Kit from Qiagen.

Manufacturer protocol

Download the product protocol from Qiagen for RNeasy Plus Mini Kit below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing RNA isolation / purification Cells - immortalized NALM-6 using RNeasy Plus Mini Kit from Qiagen. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.