RNeasy Mini Kit

RNA isolation / purification Cells - immortalized PNT1A

Experiment
RNA isolation / purification Cells - immortalized PNT1A
Product
RNeasy Mini Kit from Qiagen
Manufacturer
Qiagen

Protocol tips

Protocol tips
- To yield high RNA, give the column an extra wash with both RW1 and RPE buffers (3 total washes with each buffer).
Downstream tips
- Include DNAse treatment for 15-20min.

- Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C.

- Use water to elute the RNA that is warmed to ~60`C.

- For purifying RNA add either 10 μl β-mercapto ethanol (β-ME) or 20 μl 2 M dithiothreitol (DTT).

Publication protocol

Total RNA obtained from cultures using RNeasy mini kit (Qiagen, Hilden, Germany) was reverse transcribed (SuperScript, Invitrogen, Karlsruhe, Germany) with oligo-dT and gene specific primers for hTRPM8 (5′-cttgggcaaaacacacaatg-3′). Real-time PCR was performed on the template using the TaqMan system in an AbiPrism 7700 Sequence Detector (Applied Biosystems, Foster City, CA) or a LightCycler 480 (Roche, Mannheim, Germany). The following fragments were amplified: nt 2910–3058 from sequence NM_024080.4 was detected with the hTRPM8 probe (5′-FAM-tcgccatgtttggctacacggtgggcacc-TAMRA-3′) and nt 1632–1732 from sequence NM_003234 was detected with the hTFR probe (5′-JOE-tgaatggctagagggatacctttcgtccc-(6-TAMRA)-3′). The human transferrin receptor was used as a control template for RNA integrity and PCR performance. Relative quantification was performed using the REST software (Relative Expression Software Tool

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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?

Share your thoughts or question with experts in your field by adding a discussion!

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing RNA isolation / purification Cells - immortalized PNT1A using RNeasy Mini Kit from Qiagen.

Paper title
TRPM8 Ion Channels Differentially Modulate Proliferation and Cell Cycle Distribution of Normal and Cancer Prostate Cells
Full paper
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Manufacturer protocol

Download the product protocol from Qiagen for RNeasy Mini Kit below.

Download PDF Download manufacturer protocol

Videos

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