RNeasy Plus Mini Kit

RNA isolation / purification Cells - immortalized Caov-3

Experiment
RNA isolation / purification Cells - immortalized Caov-3
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

RNA was isolated using a Qiagen RNeasy plus mini kit (Qiagen, Hilden, Germany). Subsequently, cDNA was synthesized with random hexamer primers using the Fermentas Revertaid cDNA synthesis kit (Fermentas, Leon‐Rot, Germany). qRT‐PCR was executed with 10 ng cDNA for ICAM‐1 or VE‐cadherin expression (Taqman gene expression assay Hs00164932_m1 or Hs00174344_m1, respectively; Applied Biosystems, Carlsbad, CA) and for GAPDH expression [primers: Fw 5′‐CCACATCGCTCAGACACCAT‐3′, Rev 5′‐GCGCCCAATACGACCAAAT‐3′ and probe: CGTTGACTC CGACCTTCACCTTCCC (Eurogentec, Maastricht, The Netherlands)] using an ABI ViiA7 real‐time PCR system (Applied Biosystems). For ICAM‐4 expression detection, qRT‐PCR was performed using AbsoluteQPCR SYBRGreen‐ROXMix (Abgene, Surrey, United Kingdom) and ICAM‐4 primers: 5′‐CGGGTTGGGTGTCTTACCAG‐3′ (Fw) and 5′‐GACCGGA GGCTCCAAAATCA‐3′ (Rev). Results are shown as relative expression compared to GAPDH levels using the comparative Ct values method.

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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 Caov-3 using RNeasy Plus Mini Kit from Qiagen.

Paper title
Upregulation of endogenous ICAM-1 reduces ovarian cancer cell growth in the absence of immune cells.
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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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