RNeasy Mini Kit

RNA isolation / purification Bacteria - Gram positive Streptococcus pyogenes

Experiment

RNA isolation / purification Bacteria - Gram positive Streptococcus pyogenes

Product

RNeasy Mini Kit from Qiagen

Manufacturer

Qiagen

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Tips Publication protocol Reviews Discussion Papers Manufacturer protocol Videos

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

RNA isolation and purification were performed using an RNeasy kit (Qiagen). Purified RNA was analyzed for quality and concentration with an Agilent 2100 Bioanalyzer. cDNA was synthesized from the purified RNA using Superscript III (Invitrogen), and TaqMan quantitative RT-PCR was performed with an ABI 7500 Fast System (Applied Biosystems). Comparison of transcript levels was done using ΔCT method of analysis using tufA as the endogenous control gene

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Reviews

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Discussion

5 years ago

Author: Paul G. Macon United States

Some help with RNA isolation using Trizol

Hello! I used Trizol to extract total RNA from in-vitro cultured bacteria (1 X 10^8 cells). After phase separation, I mixed ~0.4 ml of the upper phase which contains RNA with 0.5 mL cold isopropanol. However, the amount of RNA when measured in Nanodrop was very low. In addition, the ratio between 260 and 230 was around 0.1 to 0.5. Is there a chance that my sample was contaminated by the Trizol reagent? When I collected the aqueous phase I made sure to not touch the lower phase. What should I do?

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing RNA isolation / purification Bacteria - Gram positive Streptococcus pyogenes using RNeasy Mini Kit from Qiagen.

Paper title

Crystal Structure of Peroxide Stress Regulator from Provides Functional Insights into the Mechanism of Oxidative Stress Sensing

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Manufacturer protocol

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

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Videos

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