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Real time pcr lightcycler 2

Manufactured by Roche
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The Real-Time PCR LightCycler II is a laboratory instrument used for performing real-time polymerase chain reaction (PCR) analysis. It is designed to amplify and simultaneously detect and quantify specific DNA sequences in real-time.

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Lab products found in correlation

Tri reagent, by Merck Group (10 mentions) M mlv, by Promega (8 mentions) Ex taq qpcr premix, by Roche (6 mentions) Rna rneasy plant mini kit, by Qiagen (2 mentions) Qpcr primers, by Merck Group (2 mentions) Fastgene scriptase 2 cdna kit, by Nippon Genetics (1 mentions) M mlv, by Thermo Fisher Scientific (1 mentions) Kapa sybr fast qpcr master mix, by Roche (1 mentions) Wizard sv genomic dna purification kit, by Promega (1 mentions)

Close spelling variants of this product

LightCycler 480 Real-time PCR 96-well system II QuantStudio®5 real-time PCR Design & Analysis system (LightCycler® 480 II, LightCycler 480 Instrument II Real-time PCR System machine Roche Lightcycler II real-time PCR machine LightCycler 480 II Real-Time PCR Cycler LightCycler®480 2.0 Real time PCR system LightCycler 2.0 Real-Time PCR system LightCycler 480 Instrument II real-time PCR system LightCycler® 480 II real-time qRT-PCR system LightCycler 480 II real-time PCR

14 protocols using real time pcr lightcycler 2

1

Quantitative Real-Time PCR Analysis

Cited 1 time
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Total RNA was extracted using TRI Reagent® (Sigma-Aldrich). Three μg of RNA was used for retro-transcription with M-MLV (Promega, Madison, WI). qPCR was performed in triplicates by using validated qPCR primers (BLAST), Ex TAq qPCR Premix, and the Real-Time PCR LightCycler II (Roche Diagnostics, Indianapolis, IN) as previously described40 (link). mRNA levels were normalized to actin mRNA, and the relative mRNA levels were determined by using the 2−ΔΔCt method.
Lettieri Barbato D., Tatulli G., Maria Cannata S., Bernardini S., Aquilano K, & Ciriolo M.R. (2015). Glutathione Decrement Drives Thermogenic Program In Adipose Cells. Scientific Reports, 5, 13091.
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2

Quantitative Analysis of OxPHOS Genes

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Total RNA was extracted using TRI Reagent® (Sigma-Aldrich). Three micrograms of RNA was used for retro-transcription with M-MLV (Promega, Madison, WI). qPCR was performed in triplicates by using validated qPCR primers (BLAST), Ex TAq qPCR Premix, and the Real-Time PCR LightCycler II (Roche Diagnostics, Indianapolis, IN) as previously described (Lettieri Barbato et al., 2013 (link)). mRNA levels were normalized to actin mRNA, and the relative mRNA levels were determined by using the 2−ΔΔCt method. To calculate OxPHOS gene expression ratio, nuclear-encoded OxPHOS mRNA levels were compared to mitochondrial-encoded OxPHOS mRNA. The relative mRNA levels were determined by using the 2−ΔΔCt method and were normalized to actin.
Lettieri Barbato D., Tatulli G., Vegliante R., Cannata S.M., Bernardini S., Ciriolo M.R, & Aquilano K. (2015). Dietary fat overload reprograms brown fat mitochondria. Frontiers in Physiology, 6, 272.
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3

Quantitative Gene Expression Analysis in Arabidopsis

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Total RNA was extracted from 100 mg of A. thaliana leaves using the RNeasy Plant Mini Kit RNA from Qiagen (Germantown, MD). Two micrograms of RNA were used for retro-transcription with FastGene Scriptase II cDNA-Kit (Nippon Genetics Europe, Düren, Germany). qPCR was performed in triplicates by using specific qPCR primers (Sigma-Aldrich Corporation, St. Louis, MO), the qPCRBIO SyGreen Mix (PCR Biosystem, London, UK) and the Real-Time PCR Light-Cycler II (Roche Diagnostics, Indianapolis, IN). mRNA levels were normalized to actin (ACT8), GAPDH and tubulin beta chain (TUB8) transcripts, and the relative mRNA levels were determined by using the 2-ΔΔCt method (Pfaffl, 2001 (link)). The primer sequences are listed in Supplemental Table S1.
Fiorillo A., Mattei M., Aducci P., Visconti S, & Camoni L. (2020). The Salt Tolerance Related Protein (STRP) Mediates Cold Stress Responses and Abscisic Acid Signalling in Arabidopsis thaliana. Frontiers in Plant Science, 11, 1251.
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4

RNA Extraction and qPCR Analysis

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Total RNA was extracted using TRI Reagent® (Sigma-Aldrich). RNA (3 μg) was retro-transcripted by using M-MLV (Promega, Madison, WI). qPCR was performed in triplicate by using validated qPCR primers (BLAST), Ex TAq qPCR Premix, and the Real-Time PCR LightCycler II (Roche Diagnostics, Indianapolis, IN) as previously described [14 (link)]. mRNA levels were normalized to actin mRNA, and the relative mRNA levels were determined through the 2−ΔΔCt method.
Lettieri-Barbato D., Cannata S.M., Casagrande V., Ciriolo M.R, & Aquilano K. (2018). Time-controlled fasting prevents aging-like mitochondrial changes induced by persistent dietary fat overload in skeletal muscle. PLoS ONE, 13(5), e0195912.
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5

Quantitative Real-Time PCR Analysis

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Total RNA was extracted using TRI Reagent (Sigma-Aldrich) and used for retro-transcription as previously described [48 (link)]. Three micrograms of RNA was used for retro-transcription with M-MLV (Promega). qPCR was performed in triplicates by using validated qPCR primers (BLAST), Ex TAq qPCR Premix and the Real-Time PCR LightCycler II (Roche Diagnostics, Indianapolis, IN, USA). mRNA levels were normalized to RPL mRNA, and the relative mRNA levels were determined by using the 2−ΔΔCt method. The primer sequences are listed in Table 1.
Aquilano K., Baldelli S., Barbera L.L., Barbato D.L., Tatulli G, & Ciriolo M.R. (2016). Adipose triglyceride lipase decrement affects skeletal muscle homeostasis during aging through FAs-PPARα-PGC-1α antioxidant response. Oncotarget, 7(17), 23019-23032.
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6

Quantitative RT-PCR Gene Expression Analysis

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Total RNA was extracted using the TRI reagent (Sigma-Aldrich, St. Louis, MO). Three micrograms of RNA were used for retrotranscription with M-MLV (Promega, Madison, WI). qPCR was performed in triplicates by using validated qPCR primers (nucleotide BLAST, https://blast.ncbi.nlm.nih.gov), Ex TAq qPCR Premix (Lonza Sales, Basel, Switzerland), and the Real-Time PCR LightCycler II (Roche Diagnostics, Indianapolis, IN). mRNA levels were normalized vs. actin mRNA level, and the relative mRNA levels were determined by using the 2−ΔΔCt method. The primer sequences are listed in Table 1.
Baldelli S., Limongi D., Coni C., Ciccarone F., Ciotti M., Checconi P., Palamara A.T, & Ciriolo M.R. (2021). BK Polyomavirus Activates HSF1 Stimulating Human Kidney Hek293 Cell Proliferation. Oxidative Medicine and Cellular Longevity, 2021, 9176993.
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7

RNA Extraction and qPCR Analysis

Cited 1 time
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Total RNA was extracted using TRI Reagent® (Sigma-Aldrich). RNA (3 μg) was retro-transcripted by using M-MLV (Promega, Madison, WI). qPCR was performed in triplicate by using validated qPCR primers (BLAST), Ex TAq qPCR Premix, and the Real-Time PCR LightCycler II (Roche Diagnostics, Indianapolis, IN) as described by Lettieri Barbato et al. [42 (link)]. mRNA levels were normalized to actin mRNA, and the relative mRNA levels were determined through the 2−ΔΔCt method.
Lettieri-Barbato D., D’Angelo F., Sciarretta F., Tatulli G., Tortolici F., Ciriolo M.R, & Aquilano K. (2017). Maternal high calorie diet induces mitochondrial dysfunction and senescence phenotype in subcutaneous fat of newborn mice. Oncotarget, 8(48), 83407-83418.
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8

RT-qPCR Assay for Gene Expression

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RT-qPCR analysis was carried out as previously described [22 (link)]. Briefly, total RNA was extracted using TRI reagent (Sigma-Aldrich). 3 µg of RNA was used for retrotranscription with M-MLV (Promega, Madison, WI, USA). qPCR was performed in triplicates by using validated qPCR primers (BLAST), Ex TAq qPCR Premix (Lonza Sales), and the Real Time PCR LightCycler II (Roche Diagnostics, Indianapolis, IN, USA). mRNA levels were normalized to β-actin mRNA, and the relative mRNA levels were determined by using the 2−ΔΔCt method.
Lettieri Barbato D., Tatulli G., Aquilano K, & Ciriolo M.R. (2014). Inhibition of Age-Related Cytokines Production by ATGL: A Mechanism Linked to the Anti-Inflammatory Effect of Resveratrol. Mediators of Inflammation, 2014, 917698.
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9

Quantitative Real-Time PCR Protocol

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Total RNA was extracted using TRI Reagent. Three micrograms of RNA was used for retrotranscription with M-MLV (Promega). qPCR was performed in triplicates by using validated qPCR primers (BLAST), Ex TAq qPCR Premix and the Real-Time PCR LightCycler II (Roche Diagnostics, Indianapolis, IN, USA). mRNA levels were normalized to RPL mRNA, and the relative mRNA levels were determined by using the 2−ΔΔCt method. The primer sequences are listed in Table 1.
Baldelli S., Aquilano K, & Ciriolo M.R. (2014). PGC-1α buffers ROS-mediated removal of mitochondria during myogenesis. Cell Death & Disease, 5(11), e1515-.
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10

Quantifying Arabidopsis AHA1 and AHA2 Transcripts

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Total RNA was extracted from 100 mg of Arabidopsis leaves using the RNeasy Plant Mini Kit RNA from Qiagen (Hilden, GE). Two micrograms of RNA were used for retro-transcription with M-MLV from Invitrogen (Carlsbad, CA). qPCR was performed in triplicates by using validated qPCR primers from Sigma (St. Louis, MO) specific for AHA1 and AHA2 transcripts, KAPA SYBR FAST qPCR Master mix from KapaBiosystems (Boston, MA) and the Real-Time PCR Light-Cycler II from Roche Diagnostics (Indianapolis, IN). mRNA levels were normalized to beta 8-tubulin mRNA (TUB8), and the relative mRNA levels were determined by using the 2 ÀDDCt method (Pfaffl, 2001) .
Muzi C., Camoni L., Visconti S, & Aducci P. (2016). Cold stress affects H+-ATPase and phospholipase D activity in Arabidopsis. Plant physiology and biochemistry : PPB, 108.
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