RNeasy Mini Kit

RNA isolation / purification Cells - immortalized U-251

Experiment
RNA isolation / purification Cells - immortalized U-251
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.

Publication protocol

To detect EGFR mRNA expression in glioma cells, RNA was extracted from the cell lines with RNeasy Mini Kit (Qiagen, Hilden, Germany) and quantified with NanoDrop (Thermo Fisher, Wilmington, DE). Reverse transcripts were produced using M-MLV reverse transcriptase (Invitrogen, Grand Island, NY), and PCR was conducted with GoTaq® Flexi DNA Polymerase (Promega, Madison, WI). The forward primer is TGACTCCGTCCAGTATTGATCG, and the reverse primer is GCCCTTCGCACTTCTTACACTT. The PCR reaction parameters are 95 °C 5 min, 35 cycles at 95 °C 40 s, 55 °C 40 s, 72 °C 1 min, and final extension at 72 °C for 10 min.

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Reviews

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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 U-251 using RNeasy Mini Kit from Qiagen.

Paper title
CAR-Engineered NK Cells Targeting Wild-Type EGFR and EGFRvIII Enhance Killing of Glioblastoma and Patient-Derived Glioblastoma Stem 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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