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Lightcycler480 software setup

Manufactured by Roche
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The LightCycler480 Software Setup is a software package that enables the configuration and operation of the LightCycler480 real-time PCR instrument. The software provides the necessary tools to set up, run, and analyze experiments conducted with the LightCycler480 system.

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Lab products found in correlation

Trizol reagent s5, by Sangon (2 mentions) Trizol reagent, by Sangon (2 mentions) Gallios machine, by Beckman Coulter (1 mentions)

4 protocols using lightcycler480 software setup

1

Quantifying mRNA Expression in Cell Lines

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Total cellular RNA was extracted from MCF-7 cells, L02 cells or HepG2 cells using Trizol reagent S5 (Sangon Co. Ltd., Shanghai, China) according to the indicated protocol. The cDNA samples were prepared by using the reverse transcription (RT) reaction with an AMV First Strand cDNA Synthesis Kit (BBI, Toronto, Canada). qPCR analysis of mRNA was performed with SG Fast qPCR Master Mix (2X) (BBI), according to the indicated protocol on an LightCycler 480 Software Setup (Roche). The primers used in this experiment were shown in Table S1.† We evaluated all of the data with respect to the mRNA expression by normalizing to the expression of GAPDH and using the 2–ΔΔCt method.
Huang J., Wang H., Yang X., Quan K., Yang Y., Ying L., Xie N., Ou M, & Wang K. (2016). Fluorescence resonance energy transfer-based hybridization chain reaction for in situ visualization of tumor-related mRNA †Electronic supplementary information (ESI) available. See DOI: 10.1039/c6sc00377j. Chemical Science, 7(6), 3829-3835.
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2

Quantitative RT-PCR Analysis of mRNA

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Total cellular RNA was extracted from HepG2 cells or L02 cells using Trizol reagent S5 (Sangon Co. Ltd., Shanghai, China) according to the indicated protocol. The cDNA samples were prepared by using the reverse transcription (RT) reaction with AMV First Strand cDNA Synthesis Kit (BBI, Toronto, Canada). qRT-PCR analysis of mRNA was performed with SG Fast qPCR Master Mix (2X) (BBI) on a LightCycler480 Software Setup (Roche). The primers used in this experiment were shown in Table S1.† We evaluated all the data with respect to the mRNA expression by normalizing to the expression of GAPDH and using the 2–ΔΔCt method.
Ou M., Huang J., Yang X., Quan K., Yang Y., Xie N, & Wang K. (2016). MnO2 nanosheet mediated “DD–A” FRET binary probes for sensitive detection of intracellular mRNA †Electronic supplementary information (ESI) available. See DOI: 10.1039/c6sc03162e Click here for additional data file.. Chemical Science, 8(1), 668-673.
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3

Nanoflare-Mediated miRNA Detection

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LOVE-1, 22Rv1, HeLa and SMMC-7721 cells were incubated with nanoflares. After 6 h, the cells were washed to remove the redundant particles. Cells were then detached from culture dishes using Trypsin-EDTA Solution. The solution containing treated cells was centrifuged (2000 rpm, 4 min) and resuspended in PBS three times. Flow cytometric assay was performed using Beckman Coulter Gallios machine.
Total cellular RNA was extracted from cells using Trizol reagent (Sangon Co. Ltd., Shanghai, China) according to the indicated protocol. The cDNA samples were prepared by using the reverse transcription (RT) reaction with AMV First Strand cDNA Synthesis Kit (BBI, Toronto, Canada). qPCR analysis of miRNAs were performed with SG Fast qPCR Master Mix (2X) (BBI), according to the indicated protocol on an Light Cycler480 Software Setup (Roche). The primers used in this experiment are shown in Table 1. We evaluated all the data with respect to the miRNA expression by normalizing to the expression of U6 and using the 2-∆∆Ct method.
Li J., Huang J., Yang X., Yang Y., Quan K., Xie N., Wu Y., Ma C, & Wang K. (2018). Two-Color-Based Nanoflares for Multiplexed MicroRNAs Imaging in Live Cells. Nanotheranostics, 2(1), 96-105.
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4

Quantifying miRNA-21 Expression in Cell Lines

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Total cellular RNAs were extracted from HeLa cells, HepG2 cells, MCF-7 cells, and L02 cells respectively, using Trizol reagent (Sangon Co. Ltd., Shanghai, China) according to the manufacturer's instructions. The cDNA samples were prepared by using the reverse transcription (RT) reaction with an AMV First Strand cDNA Synthesis Kit (BBI, Toronto, Canada). qPCR analysis of miRNA was performed with SG Fast qPCR Master Mix (2× BBI), according to the indicated protocol on an LightCycler480 Software Setup (Roche). The relative expression of miR-21 was calculated using the 2–ΔΔCt method.
Wei Q., Huang J., Li J., Wang J., Yang X., Liu J, & Wang K. (2018). A DNA nanowire based localized catalytic hairpin assembly reaction for microRNA imaging in live cells †Electronic supplementary information (ESI) available. See DOI: 10.1039/c8sc02943a. Chemical Science, 9(40), 7802-7808.
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