RNeasy Mini Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	To yield high RNA, give the column an extra wash with both RW1 and RPE buffers (3 total washes with each buffer).	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

Protocol tips
To yield high RNA, give the column an extra wash with both RW1 and RPE buffers (3 total washes with each buffer).
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

Total RNA was extracted from frozen PT biopsy samples using RNeasy minikits and the RNAse-Free DNAse Set (Qiagen, Germantown, MA). The RNA quantity and quality were determined with Nanodrop and Agilent analyzers. RNA from postmortem human brain tissue was used as positive control. Eight hundred micrograms of RNA were used for cDNA synthesis using the AMV First-Strand cDNA Syhthesis Kit (no. 12328-040; Invitrogen, Carlsbad, CA). Two microliters of cDNA synthesis reaction were used in a 25-μl real-time PCR reaction with FAM-labeled Taqman Gene Expression Assays for NR-2A (NMDAR2a; Hs00168219_m1) and MAP2 (Hs01103234_g1; Applied Biosystems, Foster City, CA). β-Actin (Hs03023943_g1) was used as the reference control gene. The amplification conditions were: 10 min at 95°C followed by 40 cycles of 15 s at 95°C, 1 min at 60°C. CT values were analyzed using the 2-ΔΔCT method.

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 Tissue - Human Brain using RNeasy Mini Kit from Qiagen.

Manufacturer protocol

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

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing RNA isolation / purification Tissue - Human Brain using RNeasy 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.