PureLink™ RNA Mini Kit

RNA isolation / purification Bacteria - Gram positive Staphylococcus epidermidis

Experiment

RNA isolation / purification Bacteria - Gram positive Staphylococcus epidermidis

Product

PureLink™ RNA Mini Kit from Thermo Fisher Scientific

Manufacturer

Thermo Fisher Scientific

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Tips Publication protocol Reviews Discussion Papers Manufacturer protocol Videos

Protocol tips

Upstream tips	Protocol tips	Downstream tips
For lysis of yeast, extra enzymatic or mechanical disruption is recommended. For bacteria, lysis using lysozyme is recommended	Depending on the material you want to isolate RNA from, please refer to the manual for additional reagents/lab equipment that are required	DNase is not supplied with this kit, a DNase kit is available which is compatible with this kit

Upstream tips
For lysis of yeast, extra enzymatic or mechanical disruption is recommended. For bacteria, lysis using lysozyme is recommended
Protocol tips
Depending on the material you want to isolate RNA from, please refer to the manual for additional reagents/lab equipment that are required
Downstream tips
DNase is not supplied with this kit, a DNase kit is available which is compatible with this kit

Publication protocol

RNA was isolated from NHEKs using Purelink RNA isolation columns (ThermoFisher Scientific) according to manufacturer’s instructions. RNA was quantified using a Nanodrop spectrophotometer (ThermoFisher Scientific), and 500ng of RNA was reverse-transcribed using the iScript cDNA synthesis kit (Bio-Rad, Irvine, CA). qPCR reactions were ran on a CFX96 Real-Time Detection System (Bio-Rad) using gene-specific primers and TaqMan probes (ThermoFisher Scientific).

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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

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for PureLink™ RNA Mini Kit below.

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Videos

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