TRIzol Reagent

Protocol tips

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

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

Publication protocol

SIRT1 was examined in pancreatic cancer cells and tissues before and after GEM treatment using quantitative RT-PCR (qRT-PCR). Briefly, Total RNA (1 μg) extracted (TRIzol reagent; Invitrogen, Carlsbad, CA, USA) was reverse transcribed to cDNA according to the manufacturer's instructions. Real-time PCR was carried out using the SYBR Green quantitative PCR kit (Invitrogen) according to the manufacturer's instructions using an Applied Biosystems 7300 real-time PCR system (Carlsbad, CA, USA). SIRT1 forward primer, 5′-CGGAAACAATACCTCCACCTGA-3′; reverse primer, 5′-GAAGTCTACAGCAAGGCGAGC A-3′ (amplicon size, 242 bp). All data were normalized to the GAPDH (forward primer, 5′-GACAACTTTGGCATCGT-3′; reverse primer, 5′-ATGCAGGGATGATGTTCTGG-3′; amplicon size, 133 bp). Relative SIRT1 mRNA expression was calculated using the An external file that holds a picture, illustration, etc.
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Manufacturer protocol

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

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