TRIzol Reagent

RNA isolation / purification Cells - immortalized A431

Experiment
RNA isolation / purification Cells - immortalized A431
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

The cultured A431 cells were treated with different concentrations of berberine (0, 25, 50, and 100 µg/mL), and incubated for an additional 4, 8, or 12 h. Total RNA was subsequently extracted from the cells using TRIzol reagent (Tiangen Biotech, Beijing, China). The RNA was reversetranscribed to cDNA. The cDNA samples were subjected to real-time PCR, using a Real Master Mix (SYBR Green) Kit (Tiangen Biotech). The following primers were used for PCR amplification: Bax, forward 5'-GGATGATTGCCGCCGT-3', reverse 5'-CCCAGTTGAAGTTGCCGT-3' (melting
temperature, Tm = 55°C); Bcl-2, forward 5'-TGCACCTGACGCCCTTCAC-3', reverse 5'-AGACA
GCCAGGAGAAATCAAACAG-3' (Tm = 55°C); Ezrin, forward 5'-ACGTCTGAGAATCAACAAGC-3',
reverse 5'-TTCTCCTCATAGTCCTGCAG-3' (Tm = 53°C); and GAPDH, forward 5'-GGTGAAGGTC
GGAGTCAACGGA-3', reverse 5'-GAGGGATCTCGCTCCTGGAAGA-3' (Tm = 61°C). The PCR conditions were set as follows: 1 cycle at 95°C for 10 min, 40 cycles at 95°C for 15 s and Tm for
1 min, and 72°C for 7 min. The amplification reaction, PCR product detection, and data analyses were performed using the ABI Prism7900 Sequence Detection System (Applied Biosystems,
Waltham, MA, USA)

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Discussion

Discussion

4 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 A431 using TRIzol Reagent from Thermo Fisher Scientific.

Paper title
Inhibitory effect of berberine on human skin squamous cell carcinoma A431 cells.
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Manufacturer protocol

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

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