TRIzol Reagent

RNA isolation / purification Cells - immortalized hES(h1)

Experiment
RNA isolation / purification Cells - immortalized hES(h1)
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 (Life Tech., Carlsbad CA). mRNAs were reverse-transcribed using a SuperScript III cDNA synthesis kit (Life Tech., Carlsbad CA). Gene expression of ATP8B2, DCAF12, LAT2, BIK, FNBP4, AGPAT6, AIG1, and PPTC7 in individual selected CPF resistant cells and passage matched parental cells was measured using quantitative real-time reverse transcription (RT)-PCR using primers listed in Table 4. Real-Time PCR was performed using Bio-Rad CFX96 Touchâ„¢ Real-Time PCR Detection System and a SYBR Green Supermix Kit (Bio-Rad Laboratories, Hercules, CA). The PCR efficiency was examined by serially diluting template cDNA and the melting curve data were collected to check the PCR specificity. Results were calculated using the delta, delta Ct method normalizing to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) expression for each sample.

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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

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