RNA isolation / purification Cells - immortalized KG-1

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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5 years ago

5 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 3 matching solutions for this experiment

Upstream tips
- Do not wash cells before adding reagent (suspension cells).
Protocol tips
- For low contamination rate carefully remove the upper aqueous phase.

- Adding too little reagent can lead to contamination with DNA.
Mini Total RNA Kit

IBI Scientific

Protocol tips
total RNA was isolated using a Total RNA minikit (catalog number IB147323; IBI). RNA was quantified, and equal amounts of total RNA were treated with DNase I (M6101; Promega)
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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