Cfx96 touch real time pcr detection system

Manufactured by Roche

Sourced in United States, Germany

The CFX96 Touch Real-Time PCR Detection System is a laboratory equipment designed for real-time polymerase chain reaction (PCR) analysis. It is capable of accurately detecting and quantifying nucleic acid sequences in various sample types.

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Lab products found in correlation

Trizol reagent, by Thermo Fisher Scientific (5 mentions) Taq dna polymerase, by Thermo Fisher Scientific (2 mentions) Rnasin rnase inhibitor, by Promega (2 mentions) Sybr green 1, by Roche (2 mentions) Kapa sybr fast one step qrt pcr kit, by Roche (1 mentions) Superase in rnase inhibitor, by Thermo Fisher Scientific (1 mentions) Sybr green, by Roche (1 mentions) Qiaquick spin column, by Qiagen (1 mentions) Superscript 2 rt, by Thermo Fisher Scientific (1 mentions) Dna free dna removal kit, by Thermo Fisher Scientific (1 mentions) Rneasy kit, by Qiagen (1 mentions) Transcriptor first strand cdna synthesis kit, by Roche (1 mentions) Sybr green 1 master, by Roche (1 mentions) Superscript 3 reverse transcriptase kit, by Thermo Fisher Scientific (1 mentions) Sybr green pcr master mix, by Roche (1 mentions)

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Real-time System (7 citations) CFX96 detection system (6 citations) CFX96 system (6 citations) CFX96 (6 citations)

6 protocols using cfx96 touch real time pcr detection system

Quantifying Gene Expression in Artificial Skin

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RNA from artificial skin was isolated using TRIzol Reagent (Invitrogen, USA), according to the manufacturer’s instructions. Complementary DNA synthesis and amplification were performed using the CFX96 Touch Real-time PCR Detection system thermocycler and KAPA SYBR FAST One-Step qRT-PCR Kit (KAPABIOSYSTEMS, USA), according to the manufacturers’ protocols. The primer pairs used are presented in Table 3. The level of mRNA was normalised to the levels of β-actin or TGase-1 mRNA. Subsequently, the relative mRNA expression levels were calculated using the 2^−ΔΔCt method.Table 3

Primers used for qRT-PCR analysis.

Gene	Forward	Reverse
FLG	5′-AAGGTTCACATTTATTGCCAAA-3′	5′-GGATTTGCCGAAATTCCTTT-3′
β-Actin	5′-TGACGGGGTCACCCACACTGTGCCCATCTA-3′	5′-CTAGAAGCATTTGCGGTGGACGATGGAGGG-3′
Caspase-14	5′-AAATGAGCAATCCGCGGTCTTTGG-3′	5′-CCGTGGAATAAACGTGCAAGGCAT-3′
Involucrin	5′-CTCCACCAAAGCCTCTGC-3′	5′-CTGCTTAAGCTGCT GCTC-3′
TGase-1	5′-TGAATAGTGACAAGGTGTACTGGCA-3′	5′-GTGGCCTGAGACATTGAGCAGCAT-3′
psoriasin-hS100A7	5′-AGACGTGATGACAAGATTGAC-3′	5′-TGTCCTTTTTCTCAAAGACGTC-3′
Koebnerisin 15S-hS100A15S	5′-CAAGTTCCTTCTGCTCCATCTTAG-3′	5′-AGCCTTCAGGAAATAAAGACAATC-3′
Koebnerisin 15L-hS100A15L	5′-ACGTCACTCCTGTCTCTCTTTGCT-3′	5′-TGATGAATCAACCCATTTCCTGGG-3′

Ampawong S., Kengkoom K., Sukphopetch P., Aramwit P., Muangkaew W., Kanjanapruthipong T, & Buaban T. (2020). Evaluating the effect of rice (Oryza sativa L.: SRNC05053-6-2) crude extract on psoriasis using in vitro and in vivo models. Scientific Reports, 10, 17618.

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RNA Extraction and qPCR Analysis of Phocaeicola

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Phocaeicola vulgatus ∆tdk strains were grown overnight in 5 mL TYG medium. Overnight cells were used to inoculate cultures at 10 mL TYG at a final dilution of 1:25 in biological triplicate. Cell growth was monitored and harvested at an OD₆₀₀ of ~0.4. When appropriate, cells were perturbed by 880 μM H₂O₂ and immediately harvested after 30 min of incubation at 37°C. Total RNA was extracted using a lysis buffer (Degnan et al., 2014 (link)) and prepared with a Qiagen RNeasy kit and treated with DNA-free™ DNA Removal Kit (Invitrogen).
Complementary DNA (cDNA) was generated from 500 ng total RNA using SUPERase•In™ RNase Inhibitor (Invitrogen) and Superscript-II RT (Invitrogen), where the RNA template was eventually degraded by inoculating 1 N NaOH for 30 min at 65°C and neutralized with 1 N HCl. DNA was isolated using a QIAquick spin column (Qiagen) and eluted in 10 mM Tris-Cl, pH 8.5.
Quantification of BVU3433 gene expression was measured through real-time quantitative PCR (qPCR) using Bio-Rad CFX96 Touch Real-Time PCR Detection System and SYBR Green (KAPA Biosystems) fluorescent dye. The CFX Maestro Software and ∆∆C_q method (Bookout et al., 2006 (link)) were used to process and calculate differences in BVU3433 and 16 s rRNA (Supplementary Table S2) expression. BVU3366 and BVU3378 were used as candidate genes to confirm RNA-Seq expression profiles.

Ortañez J, & Degnan P.H. (2024). Tracking and characterization of a novel conjugative transposon identified by shotgun transposon mutagenesis. Frontiers in Microbiology, 15, 1241582.

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Quantitative Real-Time PCR for Gene Expression

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Total RNA was extracted using TRIzol reagent (Invitrogen, 15596026, United States), and cDNA was synthesized with Transcriptor First Strand cDNA Synthesis kit (Roche, 04896866001, Germany). Quantitative real-time PCR was performed using SYBR Green I Master (Roche, 4707516001, Germany), and triplicate samples were run on a CFX96 Touch Real-Time PCR Detection System according to the manufacturer’s protocol. Normalised gene expression values (against GAPDH) were obtained using the ΔΔCT method. All details of the PCR primer sequences are presented in Supplemental Table S1.

Li Y., Yan J., Zhao Q., Zhang Y, & Zhang Y. (2022). ATF3 promotes ferroptosis in sorafenib-induced cardiotoxicity by suppressing Slc7a11 expression. Frontiers in Pharmacology, 13, 904314.

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Quantitative RT-PCR analysis of gene expression

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Reverse transcription reactions were conducted on total RNA extracted from ∼10 whole worms or 20 tail fragments using Trizol Reagent (Thermo Fisher) a SuperScript III Reverse Transcriptase Kit (Invitrogen). Quantitative real-time PCR was performed in biological triplicate on a Bio-Rad CFX96 Touch Real-Time PCR Detection System with SYBR Green PCR Master Mix (Roche) as per the manufacturer's instructions. Expression was normalized to control(RNAi) and the 2^−ΔΔCT method was used for relative quantification. Primer pairs for ubiquitously expressed GAPDH were used as a reference as previously described (Eisenhoffer et al., 2008 (link)). Experiments were carried out in biological and technical triplicates. All statistical analysis was carried out by performing Welch's unequal variances t-test using Microsoft Excel. *P<0.05, **P<0.01 and ***P<0.001. Graphs were generated using GraphPad Prism. All error bars indicate the s.d.

Akheralie Z., Scidmore T.J, & Pearson B.J. (2023). aristaless-like homeobox-3 is wound induced and promotes a low-Wnt environment required for planarian head regeneration. Development (Cambridge, England), 150(18), dev201777.

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Quantitative PCR Analysis of Target Genes

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Total RNA was extracted from cell lines, primary cultures or brain extracts using TRIzol reagent (Invitrogen) and complementary DNA synthesis was carried out using M-MLV reverse transcriptase in the presence of RNasin RNase inhibitor (Promega). PCR primers are all intron spanning. Quantitative real-time PCR was performed with Taq DNA polymerase (Invitrogen) and SYBR Green I (Roche) using a CFX96 Touch™ Real-Time PCR Detection System. Relative expression was calculated for each gene by the Ct method with Hprt for normalization. Sequences and expected product sizes are as follows: Crhr1 sense 5′-GGGCCATTGGGAAACTTTA-3′, antisense 5′-ATCAGCAGGACCAGGATCA-3′ (109 bp); sAC sense 5′-CCTGCATCGCTGTCTGGTAT-3′, antisense 5′-GAACTGTCGGGGTTCTTCGT-3′ (102 bp); c-fos sense 5′-ATCGGCAGAAGGGGCAAAGTAG-3′, antisense 5′-GCAACGCAGACTTCTCATCTTCAAG-3′ (172 bp); Hprt sense 5′-TGGGCTTACCTCACTGCTTTCC-3′, antisense 5′-CCTGGTTCATCATCGCTAATCACG-3′ (139 bp).

Inda C., Bonfiglio J.J., dos Santos Claro P.A., Senin S.A., Armando N.G., Deussing J.M, & Silberstein S. (2017). cAMP-dependent cell differentiation triggered by activated CRHR1 in hippocampal neuronal cells. Scientific Reports, 7, 1944.

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Quantitative Real-Time PCR for Cyclic AMP Signaling

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Total RNA was extracted from cell lines or total mouse brain extracts using TRI reagent (MRC), and complementary DNA synthesis was performed using M-MLV reverse transcription in the presence of RNasin RNase inhibitor (Promega). PCR primers were all intron spanning. cDNAs were amplified using Taq DNA polymerase (Invitrogen). Quantitative real-time PCR was performed with SYBR Green I (Roche) using a CFX96 Touch Real-Time PCR Detection System. Relative expression was calculated for each gene by the Ct method with HPRT for normalization.
Sequences and expected product sizes are as follows: sAC sense 5′-CCTGCATCGCTGTCTGGTAT-3′, antisense 5′-GAACTGTCGGGGTTCTTCGT-3′ (102 bp); AC1 sense 5′-GTGGCCAGTCGGATGGATAG-3′, antisense 5′-TTCACGCTGACTTTGCCTCT-3′ (119 bp); AC8 sense 5′-ACGTCATCATCTTCGCTTCCA-3′, antisense 5′-AGTACTCTGGGTAGGAGCAGA-3′ (150 bp); EPAC1 sense 5′-TCTTACCAGCTAGTGTTCGAGC-3′, antisense 5′-AATGCCGATATAGTCGCAGATG-3′ (223 bp); EPAC2 sense 5′-TAAAAGGCCGTTGGAGCGAT-3′, antisense 5′-GCCAGGACAGCATACCAGTT-3′ (195 bp); HPRT sense 5′-TGGGCTTACCTCACTGCTTTCC-3′, antisense 5′-CCTGGTTCATCATCGCTAATCACG-3′ (139 bp); and β-actin sense 5′-TGACGGGGTCACCCACACTGTGCCCATCTA-3′, antisense 5′-CTAGAAGCATTTGCGGTGGACGATGGAGGG-3′ (663 bp).

Inda C., dos Santos Claro P.A., Bonfiglio J.J., Senin S.A., Maccarrone G., Turck C.W, & Silberstein S. (2016). Different cAMP sources are critically involved in G protein–coupled receptor CRHR1 signaling. The Journal of Cell Biology, 214(2), 181-195.

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