TRIzol Reagent

RNA isolation / purification Cells - immortalized CMT12

Experiment
RNA isolation / purification Cells - immortalized CMT12
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 isolated from each cell line using Trizolc according to the manufacturer’s protocol and quantified using a biophotometer.d RNA was cleaned up using the RNeasy Mini kite according to the manufacturer’s protocol. RNA was reverse transcribed using Superscript II reverse transcriptasec according to the manufacturer’s protocol. PCR was carried out with the SYBR Green PCR Master Mix from PE Applied Biosystems on an ABI Prism 7700 Sequence Detection Systemf using 0.5 Amol/L of each primer and 2 μL AL cDNA template in each 25 μL AL reaction. TATA binding protein (TBP) was used as an exogenous control to compare the relative amount of target gene in different samples.

The PCR profile was as follows: 1 cycle of 95°C for 10 minutes, then 40 cycles of 95°C for 15 seconds and 60°C for 1 minute. The comparative CT method was used for relative quantitation of samples. Primers were purchased from Integrated DNA Technologies

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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 CMT12 using TRIzol Reagent from Thermo Fisher Scientific.

Paper title
Effect of Tolfenamic Acid on Canine Cancer Cell Proliferation, Specificity Protein (Sp) Transcription Factors, and Sp-Regulated Proteins in Canine Osteosarcoma, Mammary Carcinoma, and Melanoma Cells
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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for TRIzol Reagent below.

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