480 light cycler system

Manufactured by Roche

Sourced in Germany

The 480 Light Cycler System is a real-time PCR instrument designed for nucleic acid quantification and analysis. It provides a platform for sensitive and accurate detection and quantification of DNA and RNA targets.

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Lab products found in correlation

Sybr green supermix, by Thermo Fisher Scientific (1 mentions) Sybr green pcr master mix, by Thermo Fisher Scientific (1 mentions) Steponeplus instrument, by Thermo Fisher Scientific (1 mentions) Trizol, by Merck Group (1 mentions) Sybr green, by Merck Group (1 mentions) Superscript 3 reverse transcriptase, by Thermo Fisher Scientific (1 mentions) Rneasy kit, by Qiagen (1 mentions)

Close spelling variants of this product

Light Cycler II 480 System (5 citations) De Light Cycler 480 System (2 citations) Light Cycler 480 Probe Master System (9 citations) Light Cycler 480 (LC480) system (2 citations) Light Cycler 480 II detection system (47 citations) Real-Time PCR System (Light Cycler 480). (5 citations) Light Cycler 480 Instrument RT-PCR Detection System (4 citations) Light Cycler 480 Sybr System (2 citations)

4 protocols using 480 light cycler system

RNA Extraction and qPCR Analysis

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RNA was extracted from HCT116 cells using Thermo Fisher Scientific TRIzol and from liver tissue using Sigma TRI Reagent, both by following the manufacturer’s instructions. The concentration and purity of RNA were determined by absorbance at 260/280 nm.
cDNA was prepared from 1 μg of RNA using RevertAid Reverse Transcriptase for HCT116 RNA and Superscipt III for liver RNA. Oligo dT primers were used in both cDNA synthesizing reactions.
HCT116 cDNA was analyzed on a Roche 480 Light Cycler System with PerfeCTa SYBR Green SuperMix while cDNA generated from liver was analyzed on an Applied Biosystems StepOne Plus instrument with Sybr Green PCR Master Mix. HCT116 cDNA qPCR reactions were normalized internally using GAPDH while liver cDNA qPCR reactions were normalized internally using actin. Primer sequences are available in Table S3.

Haws S.A., Yu D., Ye C., Wille C.K., Nguyen L.C., Krautkramer K.A., Tomasiewicz J.L., Yang S.E., Miller B.R., Liu W.H., Igarashi K., Sridharan R., Tu B.P., Cryns V.L., Lamming D.W, & Denu J.M. (2020). Methyl-Metabolite Depletion Elicits Adaptive Responses to Support Heterochromatin Stability and Epigenetic Persistence. Molecular cell, 78(2), 210-223.e8.

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Quantitative Analysis of Gene Expression

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Total RNA isolation was performed by using trizol reagent, and the RNA was reverse-transcribed into single-stranded cDNA by using different primers corresponding to mRNA or microRNA. The Roche 480 LightCycler® system with SYBR Green dye binding to PCR product was used to quantify target mRNA or miRNA accumulation via uorescence PCR using human β-actin or U6 as a reference. Human primers of the associated genes used for quantitative PCR in this study were listed in Table 1. For the ampli cation reaction in each well, a Cp value was observed in the exponential phase of ampli cation, and the quanti cation of relative expression levels was achieved using standard curves for both target and endogenous control samples. Relative transcript abundance of a gene is expressed as 2 -⊿⊿Cp values (⊿Ct = Cp target -Cp reference ).

Tang Z.Z., Gu P.P., An X.F., Gou L.S, & Liu Y.W. (2022). Lack of miRNA-17 family mediates high glucose-induced PAR-1 upregulation in glomerular mesangial cells. Naunyn-Schmiedeberg's archives of pharmacology, 395(1).

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Quantifying AdipoR1 mRNA Levels

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The mRNA expression levels of AdipoR1 were detected in the two cell lines using RT-qPCR. Briefly, RT-qPCR was performed with SYBR Green (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) and the Light-Cycler Roche 480 system (Roche Diagnostics, Basel, Switzerland), The thermocycling conditions for qPCR were as follows: Pre-denaturation at 95°C for 30 sec, followed by 40 cycles of denaturation at 95°C for 5 sec and extension at 60°C for 30 sec. The 2^−ΔΔCq method was used to calculate relative expression and quantify (12 (link),14 (link)). The primer sequences used were as follows: AdipoR1, forward 5′-GCAGGCACATTACACGGT-3′, reverse 5′-TCCAGTTTTTTTTTTTTTTTAGAGGTC-3′; GAPDH forward 5′-CTCATGACCACAGTCCATGCC-3′, reverse 5′-GGCATGGACTGTGGTCATGAG-3′; miR-6835-3p, forward 5′-GACCCTCTGTCTTTTCACGAAAA-3′, reverse 5′-TTTTCGTGAAAAGACAGAGGGTC-3′; and U6, forward 5′-CTCGCTTCGGCAGCACA-3′ and reverse 5′-TGTGCTGCCGAAGCGAG-3′.

Wang H., Jiang L., Li Z., Wang W, & Hao C. (2018). miR-6835-3p regulates the function of pancreatic islet cells by modulating the expression of AdipoR1. International Journal of Molecular Medicine, 42(3), 1317-1326.

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RNA Extraction and RT-PCR Analysis

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Total RNA was extracted from cell lysates using RNeasy kit (Qiagen) and reverse transcribed into complementary DNA (cDNA) using SuperScript III Reverse Transcriptase (Invitrogen). RT-PCR was performed with cDNA corresponding to 50 ng total RNA using the LightCycler^® Roche 480 system (Roche) and the primers listed in Table 1 of Supplementary Data.

Solari V., Borriello L., Turcatel G., Shimada H., Sposto R., Fernandez G.E., Asgharzadeh S., Yates E.A., Turnbull J.E, & DeClerck Y.A. (2014). MYCN-dependent Expression of Sulfatase-2 Regulates Neuroblastoma Cell Survival. Cancer research, 74(21), 5999-6009.

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