RNA isolation / purification Cells - primary bovine umbilical vein endothelial cells

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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Protocol tips
The following might help to increase RNA yield :

- Heat the DEPC Water to 70°C before adding to the column.

- Increase the incubation time to 5 minutes.

- Increase the elution volume

. - Repeat the elution step with fresh DEPC Water (this may increase the yield, but decrease the concentration).

- Repeat the elution step using the eluate from the first elution (this may increase yield while maintaining elution volume).
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