TRIzol Reagent

RNA isolation / purification Cells - primary rat hippocampal neurons

Experiment
RNA isolation / purification Cells - primary rat hippocampal neurons
Product
TRIzol Reagent from Thermo Fisher Scientific
Manufacturer
Thermo Fisher Scientific

Protocol tips

Protocol tips
- For low 260/230 readings the best approach is to try washing sample (precipitation) with ethanol to desalt it.

Publication protocol

Total RNA was extracted using Trizol reagent (Invitrogen, Carlsbad, CA, USA) from neural cells of each group. The homogenate was mixed with 200 μL chloroform and centrifuged at 12 000 × g for 15 minutes, 4°C. The aqueous phase (about 0.5 mL upper layer) was precipitated with an equal volume of isopropanol and then centrifuged at 12 000 × g for 10 minutes, 4°C. After washing with 1 mL 75% (v/v) ethanol, the final RNA total pellet was resuspended in 20 μL diethyl pyrocarbonate water. Equal quantities (2.0 μg) of total RNA were converted to first-strand cDNA using Revert Aid First Stand cDNA Synthesis kit (Fermentas Life Sciences, Vilnius, Lithuania) for RT-PCR. PCR amplification was carried out using gene-specific PCR primers synthesized by GenScript Co., Ltd (Nanjing, Jiangsu Province, China).

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Discussion

Discussion

2 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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Paper title
5-Hydroxymethylfurfural from wine-processed inhibits hippocampal neuron apoptosis
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Manufacturer protocol

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