Lightcycler2

Manufactured by Bio-Rad

Sourced in Australia

The LightCycler 2.0 is a real-time PCR system designed for quantitative and qualitative nucleic acid analysis. It employs a thermal cycler with integrated optical detection capabilities to facilitate real-time monitoring of PCR amplification. The system is capable of performing various real-time PCR applications, including gene expression analysis, viral load determination, and pathogen detection.

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Lab products found in correlation

Nanodrop 1000, by Thermo Fisher Scientific (2 mentions) Reverse transcriptase kit, by Takara Bio (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Lightcycler 480 system, by Roche (1 mentions) Np 40, by Solarbio (1 mentions) Primescript rt kit, by Promega (1 mentions) Rnase inhibitor, by Promega (1 mentions) Trizol reagent, by Merck Group (1 mentions)

2 protocols using lightcycler2

Quantitative Expression Analysis of SNHG1

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Total RNA was isolated from tissues and cultured cells using Trizol reagent (Invitrogen, USA) according to the manufacturer's instructions. RNA quality and quantity were evaluated with the NanoDrop 1000 (ThermoFisher, Santa Clara, USA). The first strand cDNA was synthesized using a reverse transcriptase Kit (TaKaRa Biotechnology, Dalian, China) according to the manufacturer's guide. qRT-PCR (quantitive real-time PCR) was performed in triplicate with LightCycler2.0 (BIO-RAD) on a LightCycler 480 System (Roche Diagnostics, Thebarton, Australia). Results were normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) expression. The primers are below: SNHG1: Forward: 5'-GGGGTACCG TTCTCATTTTTCTACT GCTCGTG-3' and reverse: 5'-CGGGATCCATG TAATCAATCATTTTAT TATTTTCATC-3'. GAPDH: Forward: 5'-CGCTGAGTACGT CGTGGAGT-3'; reverse: 5'-CGTCAAAGGTGG AGGAGTGG -3'. The real-time PCRs were performed in triplicate and fold changes were calculated by the relative quantification (2 -ΔΔCt ) method.

Tang Q., Li Z., Han W., Cheng S, & Wang Y. (2020). High expression of lncRNA SNHG1 in prostate cancer patients and inhibition of SNHG1 suppresses cell proliferation and promotes apoptosis. Indian journal of pathology & microbiology, 63(4).

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Isolation and Quantification of RNA Fractions

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Nuclear and cytoplasmic fractions were isolated using 0.5% NP-40 (Solarbio) and RNase inhibitor (Promega) and total RNA was extracted using TRIzol reagent (Sigma). Thereafter, the cDNA was synthesized using the Prime Script RT kit (Promega). The RT-qPCR was performed in triplicate using LightCycler 2.0 (BIO-RAD). The values were normalized to GAPDH for the total RNA and cytoplasmic RNA, and U6 was used as an endogenous control for nuclear RNA. PCR primers were designed on NCBI (http://www.ncbi.nlm.nih.gov/tools/prim er-blast/) and the primer sequences are listed in Supplementary Table S1.

Li Y., Ren Y., Wang Y., Tan Y., Wang Q., Cai J., Zhou J., Yang C., Zhao K., Yi K., Jin W., Wang L., Liu M., Yang J., Li M, & Kang C. (2019). A Compound AC1Q3QWB Selectively Disrupts HOTAIR-Mediated Recruitment of PRC2 and Enhances Cancer Therapy of DZNep. Theranostics, 9(16), 4608-4623.

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