mirVana™ miRNA Isolation Kit, with phenol

RNA isolation / purification Cells - immortalized THP-1

Experiment
RNA isolation / purification Cells - immortalized THP-1
Product
mirVana™ miRNA Isolation Kit, with phenol from Thermo Fisher Scientific
Manufacturer
Thermo Fisher Scientific

Protocol tips

Protocol tips
- For RNA to dissolve better and to yeild high levels, preheat elution buffer at 95C.

- Addition of β-mercapto ethanol in lysis buffer can be skipped because the phenol will do the job of nuclease inactivation.

- To capture more amount of mRNA, modify the amount of lysis buffer:ethanol (from 1:1 to 1:1.5) during the binding step.
Downstream tips
- Do vortex for 2-plus minutes during Acid-Phenol:Chloroform extraction step. This will facilitate the removal of tightly bound proteins typically associated with RNA.

Publication protocol

Total RNA from PBMCs of 14 healthy donors was isolated using the RNeasy Plus Mini Kit (Qiagen, Germantown, MD, USA) according to the manufacturer’s instructions. THP-1 cells (2 × 106) were pretreated with skepinone-L (100 nM) in 6-well plates and further incubated with eLDL for 6 h. Untreated THP-1 cells were used as control. After centrifugation, the cell pellets were collected, and total RNA was isolated using the mirVana miRNA Isolation Kit (Ambion, Austin, TX, USA). Total RNA from PBMCs and THP-1 cells was reverse transcribed into cDNA with the High Capacity cDNA Reverse Transcription Kit (Thermo Fisher Scientific)

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Discussion

Discussion

5 years ago

Author: 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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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing RNA isolation / purification Cells - immortalized THP-1 using mirVana™ miRNA Isolation Kit, with phenol from Thermo Fisher Scientific.

Paper title
Selective p38α MAP kinase/MAPK14 inhibition in enzymatically modified LDL-stimulated human monocytes: implications for atherosclerosis.
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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for mirVana™ miRNA Isolation Kit, with phenol below.

Download PDF Download manufacturer protocol

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