RNA isolation / purification Cells - immortalized MDA-MB-157

When extracting nucleic acids from cell cultures, thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel.

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1 year ago

1 year ago by Ralf Friedmann Switzerland

I am having trouble using the QiageN RNAeasy mini kit for CD34+ cells. Any tips?

I am having trouble using the QiageN RNAeasy mini kit for CD34+ cells. Any tips?

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Found 4 matching solutions for this experiment

Upstream tips
- Addition of Buffer MZ depends on monolayer area, not cell number as it will reflect on DNA contamination if not suffcient.
Protocol tips
- To capture more amount of mRNA, modify the amount of lysis buffer :ethanol (from 1:1 to 1:1.5) during the binding step.
PureLink™ RNA Mini Kit

Thermo Fisher Scientific

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.
TRIzol Reagent

Thermo Fisher Scientific

Protocol tips
- For low 260/230 readings the best approach is to try washing sample (precipitation) with ethanol to desalt it.
Upstream tips
- Before beginning the ReliaPrep™ RNA Cell Miniprep System protocol, four solutions must be prepared.
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