RNA isolation / purification Cells - primary mouse dorsal root ganglion neurons

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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Upstream tips
- This kit can be used for the isolation of RNA from up to 5x10^5 cells
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.
Protocol tips
- To capture a higher amount of mRNA, modify the amount of lysis buffer :ethanol (from 1:1 to 1:1.5) during the binding step.
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