Cfx384 pcr machine

Manufactured by Bio-Rad

The CFX384 PCR machine is a real-time PCR instrument designed for high-throughput nucleic acid amplification and detection. It features a 384-well sample block and supports various qPCR applications, including gene expression analysis, genotyping, and pathogen detection.

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Lab products found in correlation

Itaq universal sybr green supermix, by Bio-Rad (2 mentions) Nanodrop 1000 spectrophotometer, by Thermo Fisher Scientific (2 mentions) Nanodrop, by Thermo Fisher Scientific (2 mentions) Superscript 3 kit, by Thermo Fisher Scientific (1 mentions) Rneasy kit, by Qiagen (1 mentions) Rt2 link first strand kit, by Qiagen (1 mentions) Rt2 sybr green pcr master mix, by Qiagen (1 mentions) Rt2 first strand kit, by Qiagen (1 mentions) Iscript advanced cdna synthesis kit, by Bio-Rad (1 mentions) Iscript cdna synthesis kit, by Bio-Rad (1 mentions) Qiazol lysis reagent, by Qiagen (1 mentions)

Close spelling variants of this product

CFX384 RT-PCR machine CFX384 real-time PCR machine CFX384 Touch Real-Time PCR machine CFX384 real-time system qRT-PCR machine CFX384 qRT-PCR Machine CFX384 machine PCR machine CFX384

5 protocols using cfx384 pcr machine

Comprehensive RNA Expression Analysis

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Total RNA was isolated by an RNeasy kit (#74104, Qiagen, Valencia, CA) and reverse transcribed into cDNA by a Superscript III kit (#11752, Invitrogen, CA). Quantitative PCR reactions were run with Fast Universal PCR master mix (#4352042, Invitrogen) in an ABI Prism 7700 Fast sequence detector System²⁷. The mRNA expression was determined by using cataloged primers (Invitrogen) for human FPRL1 (Hs02759175_s1) and collagen COL1A2 (Hs01028956_m1).
For PCR arrays, RNA was converted to cDNA with RT^{2 (link)} First Strand kit (#330401, Qiagen). Mouse antibacterial response PCR arrays (PAMM-148Z) and human fibrosis PCR arrays (PAHS-120Z) were performed with RT^{2 (link)} SYBR Green PCR master mix (#330501, Qiagen) in Bio-Rad CFX384 PCR machine. All gene expression analyses were normalized to 18 S (Hs99999901_s1). Results were expressed as relative fold difference.

Xu C., Ghali S., Wang J., Shih D.Q., Ortiz C., Mussatto C.C., Lee E.C., Tran D.H., Jacobs J.P., Lagishetty V., Fleshner P., Robbins L., Vu M., Hing T.C., McGovern D.P, & Koon H.W. (2017). CSA13 inhibits colitis-associated intestinal fibrosis via a formyl peptide receptor like-1 mediated HMG-CoA reductase pathway. Scientific Reports, 7, 16351.

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Quantitative Real-Time RT-PCR for Antibacterial Response

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Quantitative real-time RT-PCR was performed as previously described ^{10 (link)}. For PCR arrays, RNA was converted to cDNA with RT² First Strand kit (#330401, Qiagen). Mouse antibacterial response PCR arrays (PAMM-148Z) were performed with the RT² SYBR Green PCR master mix (#330501, Qiagen) in Bio-Rad CFX384 PCR machine. Results were expressed as relative fold difference.

Wang J., Ghali S., Xu C., Mussatto C.C., Ortiz C., Lee E.C., Tran D.H., Jacobs J.P., Lagishetty V., Faull K.F., Moller T., Rossetti M., Chen X, & Koon H.W. (2018). Ceragenin CSA13 Reduces Clostridium difficile Infection in Mice by Modulating the Intestinal Microbiome and Metabolites. Gastroenterology, 154(6), 1737-1750.

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TaqMan Genotyping of SNPs

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TaqMan assay design was made through the assay design pipeline of Life Technologies for rs35201073 and rs11577368 and TaqMan genotyping was performed according to the manufacturer’s instructions on a CFX384 PCR machine (Bio-Rad Laboratories Inc.).

Henmyr V., Lind-Halldén C., Halldén C., Säll T., Carlberg D., Bachert C, & Cardell L.O. (2016). Chronic Rhinosinusitis Patients Show Accumulation of Genetic Variants in PARS2. PLoS ONE, 11(6), e0158202.

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Quantitative Gene Expression Analysis

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Total RNA isolation was carried out using TRIsure^TM reagent following the manufacturer's instructions (Bioline Reagent). The liver, hippocampus, or cortex samples were homogenized using TissueLyser LT sample disruption apparatus (QIAGEN). RNA quantity and purity were measured with NanoDrop™ 1000 Spectrophotometer (Thermo Scientific). RNA retrotranscription was done using iScript™ Advanced cDNA Synthesis Kit (Bio‐rad). Gene expression was analyzed by real‐time quantitative PCR (RT‐qPCR) on a Bio‐Rad CFX‐384 PCR machine at the Analysis and Photodocumentation Service of the Universitat Autonòma de Barcelona. Each reaction contained 25 ng of cDNA, 7.5 μl of iTaqTM Universal SYBR Green Supermix (Bio‐Rad), and a primer concentration of 0.2 nM, with a final reaction volume of 15 μl. Primers used are listed in Table S2.
The analysis of qPCR data was done following the ΔΔCt method. Cycle threshold (Ct) were normalized subtracting from each experimental Ct, the difference of its housekeeping (HK) Ct in regard to the HK’s average value. HK genes used were β‐actin and GAPDH. Melting curves were also analyzed to ensure unique amplificon generation. Each sample (n = 6 per group) was tested at least in duplicates, and Cts higher than 37 cycles were considered as not amplified.

Roig‐Soriano J., Griñán‐Ferré C., Espinosa‐Parrilla J.F., Abraham C.R., Bosch A., Pallàs M, & Chillón M. (2022). AAV‐mediated expression of secreted and transmembrane αKlotho isoforms rescues relevant aging hallmarks in senescent SAMP8 mice. Aging Cell, 21(4), e13581.

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Quantification of Secreted Klotho mRNA

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Total RNA was extracted from the tissue samples using QIAzol Lysis Reagent (Qiagen, Madrid, Spain), then quantified on a NanoDrop spectrophotometer (Thermo Scientific, Madrid, Spain), and reverse transcribed into complimentary DNA (cDNA) with iScript cDNA Synthesis Kit (Bio-Rad, El Prat de Llobregat, Spain). Gene-specific primers used for the quantitative PCR (qPCR) analysis of messenger RNA (mRNA) s-KL levels were: s-KL Fwd: 5′-TGGCTTTCCTCCTTTACCTG-3′; s-KL Rv: 5′-GCCGACACTGGGTTTTGT-3′; m36B4 Fwd: 5′-ATGGGTACAAGCGCGTCCTG-3′; m36B4 Rv: 5′-AGCCGCAAA TGCAGATGGATC-3′; CMV Fwd: 5′-TCCCGGTGTCTTCTATGGAGG-3′; CMV Rv: 5′-CAACTCCGCCCCATTGACGCA-3′. Quantitative qPCR was performed as previously described by Masso et al., 11 on a Bio-Rad CFX-384 PCR machine at the Analysis and Photodocumentation Service of the Universitat Autonoma Barcelona using iTaqTM Universal SYBR Green Supermix (Bio-Rad). The analysis of qPCR output data followed the manufacturersuggested ΔCt method. Cycle thresholds (Ct) were measured, and the relative expression of genes was calculated by comparison of Ct values. Melt-curve analysis was used to confirm the production of a single amplicon for each gene tested. On the basis of the RT-qPCR assay efficiency, gene amplification at a level higher than 35 cycles was considered to have no expression. A 'no template control' was also included in each run.

Massó A., Sánchez A., Bosch A., Giménez-Llort L, & Chillón M. (2018). Secreted αKlotho isoform protects against age-dependent memory deficits. Molecular psychiatry, 23(9).

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