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7500 fast sequence detection software v1

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The 7500 Fast sequence detection software v1.3.1 is a software application designed for use with the 7500 Fast Real-Time PCR System. The core function of this software is to enable data acquisition, analysis, and reporting capabilities for real-time PCR experiments conducted on the 7500 Fast platform.

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Lab products found in correlation

Abi prism 7500 fast real time pcr system, by Thermo Fisher Scientific (3 mentions) Prism 7500 fast real time pcr system, by Thermo Fisher Scientific (2 mentions) High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (2 mentions) 7500 fast real time pcr system, by Thermo Fisher Scientific (2 mentions) Lc faststart dna master sybr green 1 kit, by Roche (1 mentions)

7 protocols using 7500 fast sequence detection software v1

1

Quantitative PCR Analysis of Gene Expression

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Total RNA from endothelial cells or aorta tissue from mice were isolated using Trizol reagent according to the manufacturer’s protocol. cDNA was synthesized using a High Capacity cDNA reverse transcription kit [28 (link),51 (link)]. The expressions of IL-1 beta, IL-6, TNF-alfa and MCP-1 in endothelial cells and the expression of endothelial nitric oxide synthase (Nos3), inducible nitric oxide synthase (Nos2), Nrf-2 factor (Nfe2l2), glutathione peroxidase-1 (Gpx1) and superoxide dismutase-2 Mn (Sod2-Mn) in aortas from mice were determined by quantitative PCR (ABI Prism 7500 Fast Real-Time PCR System) and analyzed with 7500 Fast sequence detection software v1.3.1 (Applied Biosystems Inc., Foster City, CA, USA), using specific TaqMan assays and Double delta Ct method. TaqMan probes used for mice were Nos3 (Mm00435217_m1), Nos2 (Mm00440502_m1), Nfe2l2 (Mm00477784_m1), Gpx1 (Mm00656767_g1), Sod2-Mn (Mm01313000_m1) and Actb (Mm01205647_g1), and for human cells they were IL1B (Hs01555410_m1), IL6 (Hs00174131_m1), CCL2 (Hs00234140_m1), TNF-alfa (Hs00174128_m1) and ACTB (Hs99999903_m1).
Asenjo-Bueno A., Alcalde-Estévez E., El Assar M., Olmos G., Plaza P., Sosa P., Martínez-Miguel P., Ruiz-Torres M.P, & López-Ongil S. (2021). Hyperphosphatemia-Induced Oxidant/Antioxidant Imbalance Impairs Vascular Relaxation and Induces Inflammation and Fibrosis in Old Mice. Antioxidants, 10(8), 1308.
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2

Quantitative Analysis of Adipogenesis Genes

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Total RNA was isolated from C2C12 cells using TRIzol reagent according to the manufacturer’s protocol. cDNA was synthesised using a High-Capacity cDNA Reverse Transcription Kit. Gene expression was measured through quantitative PCR (ABI Prism 7500 Fast Real-Time PCR System) and analysed using 7500 Fast Sequence Detection Software v.1.3.1 (Applied Biosystems Inc., Foster City, CA, USA) using specific mouse TaqMan genes and Double delta Ct method. TaqMan genes: FABP4 (Mm00445878_m1), PPAR-γ (Mm00440940_m1), and the endogenous control GAPDH (Mm99999915_g1).
Alcalde-Estévez E., Sosa P., Asenjo-Bueno A., Plaza P., Olmos G., Naves-Díaz M., Rodríguez-Puyol D., López-Ongil S, & Ruiz-Torres M.P. (2021). Uraemic toxins impair skeletal muscle regeneration by inhibiting myoblast proliferation, reducing myogenic differentiation, and promoting muscular fibrosis. Scientific Reports, 11, 512.
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3

Quantitative Analysis of Endothelin System Gene Expression

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Total RNA from EA cells or aorta tissue from rats was isolated using TRIzol reagents according to the manufacturer's protocol. cDNA was synthesized using a High‐Capacity cDNA reverse transcription kit (Martínez‐Miguel et al., 2009), and ECE‐1, prepro‐endothelin‐1 (ppET‐1), and beta‐actin or GAPDH expression were measured by quantitative PCR (ABI Prism 7 500 Fast Real‐Time PCR System) and analyzed with 7 500 Fast sequence detection software v1.3.1 (Applied Biosystems Inc., Foster City, CA, USA), using TaqMan genes and Double delta Ct method. TaqMan genes: ECE‐1 (Hs01043735_m1, Rn00585943_m1), prepro‐ET‐1 (Hs00174961_m1) and the endogenous control beta‐actin (Hs99999903_m1) and GAPDH (Rn01775763_g1) were used.
Olmos G., Martínez‐Miguel P., Alcalde‐Estevez E., Medrano D., Sosa P., Rodríguez‐Mañas L., Naves‐Diaz M., Rodríguez‐Puyol D., Ruiz‐Torres M.P, & López‐Ongil S. (2017). Hyperphosphatemia induces senescence in human endothelial cells by increasing endothelin‐1 production. Aging Cell, 16(6), 1300-1312.
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4

Quantifying Gene Expression in C2C12 Cells

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Total RNA from C2C12 cells was isolated using TRIzol reagents according to the manufacturer’s protocol. cDNA was synthesized using a High-Capacity cDNA reverse transcription kit (Applied Biosystems Inc., Foster City, CA, USA), the gene expression was measured by quantitative PCR (ABI Prism 7500 Fast Real-Time PCR System) and analyzed with 7500 Fast sequence detection software v1.3.1 (Applied Biosystems Inc., Foster City, CA, USA), using TaqMan and SYBR green genes and Double delta Ct method. TaqMan genes: MCP1 (Mm00441242_m1) and the endogenous control beta-actin (Mm00607939_s1). SYBR green primers: IL6 (from 5 to 3), forward: CCG GAG AGG AGA CTT CAC AGA GGA, and reverse: AGC CTC CGA CTT GTG AAG TGG TAT A; TNFα (from 5 to 3), forward: TGG CCC AGA CCC TCA CAC TCA, and reverse: GGC TCA GCC ACT CCA GCT GC and the endogenous control glyceraldehyde-6-phosphate dehydrogenase (GAPDH) (from 5 to 3), forward: CCA CCC AGA AGA CTG TGG AT, and reverse: CACATT GGG GGT AGG AAC AC.
Sosa P., Alcalde-Estevez E., Plaza P., Troyano N., Alonso C., Martínez-Arias L., Evelem de Melo Aroeira A., Rodriguez-Puyol D., Olmos G., López-Ongil S, & Ruíz-Torres M.P. (2018). Hyperphosphatemia Promotes Senescence of Myoblasts by Impairing Autophagy Through Ilk Overexpression, A Possible Mechanism Involved in Sarcopenia. Aging and Disease, 9(5), 769-784.
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5

Quantifying Muscle Atrophy Gene Expression

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Total RNA from quadriceps muscles were isolated using TRIzol reagent according to the manufacturer's protocol. cDNA was synthesized using a High‐Capacity cDNA reverse transcription kit. The expression of atrogin‐1 and glyceraldehyde 3‐phosphate dehydrogenase (GAPDH) was measured by quantitative PCR using Taqman genes: atrogin‐1 (Mm00499523_m1) and the endogenous control GAPDH (Mm99999915_g1). The ABI Prism 7500 Fast Real‐Time PCR System was used and analysed with 7500 Fast sequence detection software v1.3.1 (Applied Biosystems Inc., Foster City, CA, USA) with the double delta Ct method.
Alcalde‐Estévez E., Sosa P., Asenjo‐Bueno A., Plaza P., Valenzuela P.L., Naves‐Díaz M., Olmos G., López‐Ongil S, & Ruiz‐Torres M.P. (2023). Dietary phosphate restriction prevents the appearance of sarcopenia signs in old mice. Journal of Cachexia, Sarcopenia and Muscle, 14(2), 1060-1074.
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6

Quantitative RT-PCR of EA Cells

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Total cellular RNA extracted from EA cells was reverse transcribed with high capacity cDNA reverse transcription kit. 30, 39 ECE-1, prepro-endothelin-1 (ppET-1), and beta-actin complementary DNA were amplified by quantitative polymerase chain reaction by using TaqMan gene expression assay on 96-well plates in duplicate using the 7500 Fast Real-Time PCR System and analysed with 7500 Fast sequence detection software v1.3.1 (Applied Biosystems, CA, USA).
Martínez-Miguel P., Medrano-Andrés D., Griera-Merino M., Ortiz A., Rodríguez-Puyol M., Rodríguez-Puyol D, & López-Ongil S. (2017). Tweak up-regulates endothelin-1 system in mouse and human endothelial cells. Cardiovascular research, 113(2).
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7

Quantitative PCR Analysis of β-actin and GAPDH

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Endogenous β-actin and GAPDH genes were amplified by qPCR with LC FastStart DNA Master SYBR Green I Kit (Roche, Mannhein, Germany), in direct DNA (250ng in each reaction, carried out in triplicate) samples. The pairs of primers used were: human GAPDH: 5´-TCC ACT GGC GTC TTC ACC-3´ (forward) and human GAPDH: 5´-GGC AGA GAT GAC CCT TTT-3´ (reverse); human β-actin: 5´-TCA CCC ACA CTG TGC CCA TCT ACG A-3´ (forward) and human β-actin: 5´-CAG CGG AAC CGC TCA TTG CCA ATG G-3´(reverse). qPCR was performed employing the 7500 Fast real-time PCR System and 7500 Fast sequence detection software v1.3.1 (Applied Biosystems, CA, USA) as per the following cycling conditions: 50ºC for 2min and 95ºC for 10 min, followed by 45 cycles of 95ºC for 15 sec, and 60ºC for 1min.
6 Analysis of total RNA extraction and RNA purity, integrity and functionality.
Calleros-Basilio L., Cortés M.A., García-Jerez A., Luengo-Rodríguez A., Orozco-Agudo A., Valdivielso J.M., Rodríguez-Puyol D, & Rodríguez-Puyol M. (2016). Quality Assurance of Samples and Processes in the Spanish Renal Research Network (REDinREN) Biobank. Biopreservation and biobanking, 14(6).
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