RNA isolation / purification Cells - immortalized Ku812

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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Upstream tips
- This kit can be used for the isolation of RNA from up to 5x10^5 cells, or up to 5 mg of tissue.
Downstream tips
- Always blank the spectrophotometer with your elution buffer.

- When measuring 260 nm absorbance, a unit of 1 on this scale equals to 40 ng of RNA.
TRI Reagentâ„¢ Solution

Thermo Fisher Scientific

Upstream tips
- 1-Bromo-3-chloropropane is less toxic than chloroform and its use for phase separation decreases the possibility of contaminating RNA with DNA.

- The chloroform used for phase separation should not contain isoamyl alcohol or other additives.
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