RNeasy Mini Kit

RNA isolation / purification Cells - immortalized MIA PaCa-2

Experiment
RNA isolation / purification Cells - immortalized MIA PaCa-2
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 was isolated from the indicated cells using RNeasy Mini Kit (Qiagen, Valencia, CA, USA) according to the manufacturer's protocol. Reverse transcription was performed with 1 μg of total RNA using SuperScript II reverse transcriptase (Life Technologies) according to the manufacturer's instructions. The synthesized complementary DNA was amplified by PCR using specific primer sets. The PCR products were visualized by electrophoresis on agarose gels with ethidium bromide staining. GAPDH served as an internal control.

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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 MIA PaCa-2 using RNeasy Mini Kit from Qiagen.

Paper title
Assessment of the stromal contribution to Sonic Hedgehog‐dependent pancreatic adenocarcinoma
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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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