RNA isolation / purification Cells - immortalized MA-104

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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4 years ago

4 years 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

TRI Reagent® MRC

Molecular Research Center, Inc.

Upstream tips
- Avoid washing cells before the addition of TRI Reagent as this may contribute to RNA degradation.
Protocol tips
- Perform centrifugation for phase separation in the cold ( 4 - 10 C ). If performed at elevated temperatures, a
residual amount of DNA may sequester in the aqueous phase.
Upstream tips
- To wipe off
RNase, the glassware can be roasted at 150℃ for 4 hours,
while plastic can be dipped in 0.5 M NaOH for 10min, washed
by RNase-Free ddH2O thoroughly, and sterilized
TRIzol™ LS Reagent

Thermo Fisher Scientific

Upstream tips
- Permits larger samples to be processed.
Protocol tips
- Allows to perform sequential precipitation of RNA, DNA, and proteins from a single sample.
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.
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