RNA isolation / purification Cells - immortalized H1299

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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2 years ago

2 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

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.
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
Absolutely RNA Microprep Kit

Agilent Technologies

Protocol tips
- Additional vortexing, pipetting, and/or by increasing the volume of Lysis Buffer–β-ME mixture to cells at step-2 can decrease DNA contamination,
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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