Real time pcr system software

Manufactured by Thermo Fisher Scientific

Sourced in Singapore

The Real-Time PCR System Software is a computational tool used to analyze and interpret data generated from real-time polymerase chain reaction (PCR) experiments. The software provides essential functionalities for experimental setup, data acquisition, and analysis, enabling researchers to monitor and quantify target nucleic acid sequences in real-time.

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Lab products found in correlation

3 protocols using real time pcr system software

Quantitative Gene Expression Analysis of Heart and Macrophage

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Total RNA from heart tissues (left ventricles) and BMDMs were isolated using TRIzol (Life Technologies, Singapore, catalog no. 15596018) and Favorgen RNA extraction kit (Favorgen Biotech, catalog no. FATRK 001–2), respectively. The concentration and quality of RNA were measured with UV spectrophotometry (NanoDrop Technologies, Wilmington, North Carolina, USA). For cDNA synthesis, total RNA was reverse transcribed using random hexamers and SuperScript III FirstStrand Synthesis system (Life Technologies, catalog no. 18080–051). Gene expression was performed by quantitative RT-PCR (ABI PRISM 7900 or ViiA7 Real-Time PCR System) using 10-μl reaction mixture containing 20 ng cDNA, SYBR Green Master Mix (Life Technologies, catalog no. 4368702), 6-μM forward primer, and 6-μM reverse primer. Results were analyzed with Real-Time PCR System Software (Applied Biosystems, Singapore). All mRNA data were normalized against the reference gene Gapdh.

Mia M.M., Cibi D.M., Abdul Ghani S.A., Song W., Tee N., Ghosh S., Mao J., Olson E.N, & Singh M.K. (2020). YAP/TAZ deficiency reprograms macrophage phenotype and improves infarct healing and cardiac function after myocardial infarction. PLoS Biology, 18(12), e3000941.

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Quantitative RT-PCR Analysis of Osteoclastogenic Gene Expression

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Total RNA was extracted from BMM cells treated with different concentrations of HxTME at two different time points (3 and 7 days) using TRIzol reagent (Invitrogen Life Technologies, NY, USA) and reverse transcribed to cDNA using superscript cDNA synthesis kit (Invitrogen Life Technologies, NY, USA) as per manufactures instructions. The cDNA was used to quantitate different target genes using RT-PCR. Quantitative RT-PCR for different genes was performed with PRISM 7700 (PE Applied Biosystems, Foster City, CA). Samples were analyzed using TaqMan primer sets purchased from Applied Biosystems: NFATc, Mm00479445_m1; Fos, Mm00487425_m1; Cathepsin K, Mm00484036_m1; Trap, Mm00446003_m1; and 18sRNA, Mm03928990_g1. All values were normalized to the expression of the housekeeping gene 18s RNA. Relative gene expression was quantified by using Applied Biosystems real time PCR system software.

Chaugule S., Kashipathi Sureshbabu S., Dakave S., Krishna C.M., Chaudhari P., Indap M, & Chiplunkar S. (2019). Hexane Fraction of Turbo brunneus Inhibits Intermediates of RANK-RANKL Signaling Pathway and Prevent Ovariectomy Induced Bone Loss. Frontiers in Endocrinology, 10, 608.

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Quantitative RT-PCR Assay Protocol

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PCR reactions were performed in a 20 μL reaction volume in a 96-well plate (Applied Biosystems, United States). The PCR mixture contained 2 μL of cDNA template, 10 μL of commercial (2 ×) Power SYBR^® Green PCR Master Mix (Applied Biosystems), and each primer at a final concentration of 0.5 μmol/L. Real-time PCR runs were carried out using an ABI Thermal cycler 7500 real-time PCR instrument (Applied Biosystems). The conditions for thermal cycling were as follows: initial denaturation at 95 °C for 10 min, followed by 40 amplification cycles at 95 °C for 15 s and then 60 °C for 1 min. For each PCR run, a negative (no-template) control was used to test for false-positive results or contamination. The absence of nonspecific amplification was confirmed by generating a melt curve using the Applied Biosystems real-time PCR system software. The primers utilized in this study are summarized in Table 1.

Kunanopparat A., Kimkong I., Palaga T., Tangkijvanich P., Sirichindakul B, & Hirankarn N. (2016). Increased ATG5-ATG12 in hepatitis B virus-associated hepatocellular carcinoma and their role in apoptosis. World Journal of Gastroenterology, 22(37), 8361-8374.

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