RNeasy Plus Mini Kit

RNA isolation / purification Cells - immortalized Human blood cord

Experiment
RNA isolation / purification Cells - immortalized Human blood cord
Product
RNeasy Plus Mini Kit from Qiagen
Manufacturer
Qiagen

Protocol tips

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

Total RNA was isolated from cells using TRIzol reagent (Invitrogen) and RNeasy Plus Mini Kit (Qiagen). Genomic DNA was eliminated with RNase free DNase 1 (Roche), and total RNA was then subjected to another round of purification by phenol solution with pH4.3 ± 0.2 (Sigma-Aldrich). For pluripotent gene expression analysis, cDNA was generated by SuperScript II reverse transcriptase (Invitrogen) with random hexamers (Invitrogen) from 1 µg of total RNA, and real-time PCR was performed with GoTaq qPCR Master Mix (Promega).

For Gdf3 RNA was isolated from whole bone marrow mononuclear cells or CD45.2+ / LSK cells using TRIzol reagent (Invitrogen) and RNeasy mini kit (QIAGEN). cDNA was synthesized with Superscript First-Strand Synthesis System according to manufacturer's instructions (Invitrogen). qPCR was performed with SYBR reagent (Bio-Rad).

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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

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Manufacturer protocol

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

Download PDF Download manufacturer protocol

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