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7500 sds real time pcr system

Manufactured by Thermo Fisher Scientific
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The 7500 SDS Real-Time PCR System is a laboratory instrument designed for conducting real-time polymerase chain reaction (PCR) analysis. It is capable of detecting and quantifying specific DNA or RNA sequences in samples. The system provides accurate and reliable results for various applications in life science research and molecular diagnostics.

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Lab products found in correlation

Rneasy kit, by Qiagen (2 mentions) Rneasy micro kit, by Qiagen (1 mentions) Rnalater tissue storage reagent, by Merck Group (1 mentions) Lightcycler 480 real time pcr system, by Roche (1 mentions) Sybr green pcr master mix, by Thermo Fisher Scientific (1 mentions) Quantitect reverse transcription kit, by Qiagen (1 mentions) Faststart taqman probe master mix, by Roche (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Fam labeled probe, by Thermo Fisher Scientific (1 mentions) Ppargc1a, by Thermo Fisher Scientific (1 mentions) Ppargc1b, by Thermo Fisher Scientific (1 mentions) High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (1 mentions) Sybr select master mix, by Thermo Fisher Scientific (1 mentions) Qiaamp dna micro kit, by Qiagen (1 mentions)

Spelling variants of this product

7500 system (124 citations) 7500 Real-Time PCR (79 citations) 7500 PCR system (66 citations) Real-Time System (23 citations) SDS 7500 FAST Real-Time PCR system (3 citations) 7500 Real-Time PCR System SDS Software (2 citations) ABI 7500 Real-time PCR System SDS Software (2 citations)

6 protocols using 7500 sds real time pcr system

1

DNA Mixture Analysis Protocol

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The Cuyahoga County Regional Forensic Science Laboratory (CCRFSL) prepared 18 DNA mixture samples. The number of contributors ranged from 2 through 10, with two samples per contributor number. To design the DNA mixtures, reference genotypes for a sample were randomly drawn from 20 preset male and female individuals of predominantly Caucasian descent. Mixture weights were randomly drawn from a uniform Dirichlet distribution; these designed mixture proportions are shown in Table 1.
CCRFSL generated the STR mixture data in their laboratory. DNA quantity was measured with Applied Biosystems® (AB, Foster City, CA) Quantifier® Duo quantification kit using a 7500 SDS Real‐Time PCR System. Each 0.5 ng DNA mixture sample was amplified using the PowerPlex® Fusion amplification system (Promega, Madison, WI).
A 0.5 μL volume of amplified DNA product was size‐separated using an AB 3500 genetic analyzer. The analyzer recorded electropherogram (EPG) data as .hid files. CCRFSL determined a ratio of 0.37 for 3130 to 3500 AB genetic analyzer relative fluorescence units (RFU). TrueAllele analysis multiplied peak heights by this ratio, rescaling 3500 data down to the 3130 levels specified in prior probability parameters.
Bauer D.W., Butt N., Hornyak J.M, & Perlin M.W. (2019). Validating TrueAllele® Interpretation of DNA Mixtures Containing up to Ten Unknown Contributors. Journal of Forensic Sciences, 65(2), 380-398.
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2

Quantitative RT-PCR Analysis of RNA

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Total RNA was extracted from 500,000 cells using the Qiagen RNeasy kit (#74104) according to manufacturer’s protocol. RNA quality was assessed using the Nanodrop and 550 ng RNA was used to create cDNA using the TaqMan Reverse Transcription kit (N808-0234). 10 ng cDNA was used in duplicate qPCR reactions using master mix from TaqMan Universal PCR Master Mix (#4324018) and TaqMan primer probes. The following primer probes that spans exons were used to quantify mRNA levels: RHOA #HS00236938_M1, HIF-1α, #HS00153153_M1, HIF-2α #HS01026149_M1. qPCR was performed on an Applied Biosystems 7500 SDS real time PCR system. GAPDH primer probes were used for normalization. Relative expression was calculated using the delta Ct method.
Magdaleno C., Dixon L., Rajasekaran N, & Varadaraj A. (2020). HIFα independent mechanisms in renal carcinoma cells modulate divergent outcomes in fibronectin assembly mediated by hypoxia and CoCl2. Scientific Reports, 10, 18560.
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3

Quantifying HIF-1α mRNA Levels

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Total RNA was extracted from NKL cells using the Qiagen RNeasy kit (#74104) according to manufacturer’s protocol. RNA quality was assessed using the Nanodrop, and 550 ng RNA was used to create cDNA using the TaqMan Reverse Transcription kit (N808-0234). Ten nanograms of cDNA was used in duplicate qPCRs using master mix from TaqMan Universal PCR Master Mix (#4324018) and TaqMan primer probes. The HIF-1α, #HS00153153_M1 primer probes that span exons were used to quantify mRNA levels. qPCR was performed on an Applied Biosystems 7500 SDS real-time PCR system. β-Actin primer probes were used for normalization. Relative expression was calculated using the delta Ct method.
Cluff E., Magdaleno C.C., Fernandez E., House T., Swaminathan S., Varadaraj A, & Rajasekaran N. (2022). Hypoxia-inducible factor-1 alpha expression is induced by IL-2 via the PI3K/mTOR pathway in hypoxic NK cells and supports effector functions in NKL cells and ex vivo expanded NK cells. Cancer Immunology, Immunotherapy, 71(8), 1989-2005.
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4

RNA Extraction and Real-Time PCR Analysis

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Mesenteric arteries were dissected in ice cold PBS and stored at -20°C in RNAlater Tissue Storage Reagent (Sigma-Aldrich). RNA extraction was performed using the RNeasy micro kit (Qiagen, Hilden, Germany). A quantity of 100 ng of total RNA were subjected to reverse transcription with the QuantiTect Reverse Transcription kit (Qiagen). Real-time PCR was performed with Sybr Green PCR master mix (Applied Biosystems, Foster, CA, USA) on using a LightCycler 480 Real-Time PCR System (Roche, Branchburg, NJ, USA). 7500 SDS Real-Time PCR system (Applied Biosystems). Relative expression was calculated according to the formula E = 2 -(Ct[Target] -Ct[Reference]) . All data were normalized to the Gusb, Hprt and Gapdh mRNA Gapdh, Hprt and Gusb were used as housekeeping genes. Analysis was not performed when Ct values exceeded 35. Results were expressed as: 2(Ct target-Ct housekeeping gene).
Robert P., Nguyen P.M.C., Richard A., Grenier C., Chevrollier A., Munier M., Grimaud L., Proux C., Champin T., Lelièvre E., Sarzi E., Vessières E., Henni S., Prunier D., Reynier P., Lenaers G., Fassot C., Henrion D, & Loufrani L. (2021). Protective role of the mitochondrial fusion protein OPA1 in hypertension. FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 35(7).
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5

Quantitative PCR Analysis of Muscle Genes

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Total RNA was isolated from quadriceps and LA/BC muscles with TRIzol Reagent (Life Technologies) according to the manufacturer’s instructions. RNA (1 μg) was reverse transcribed to cDNA using the High Capacity cDNA Reverse Transcription Kit (Applied Biosystems). Gene-specific primers with FAM-labeled probes were from Applied Biosystems: Nr4a1, Mm01300401_m1; Prkar2a, Mm00435916_m1; Prkag3, Mm00463997_m1; Hk2, Mm00443385_m1; Slc2a4, Mm01245502_m1; Phkg1, Mm02580948_m1; Pygm, Mm00478582_m1; Ppargc1a, Mm01208835_m1; Ppargc1b, Mm00504720_m1; Bbc3, Mm00519268_m1. AR-specific forward primer 5’-CCAGTCCCAATTGTGTCAAA-3’ and reverse primer 5’-TCCCTGGTACTGTCCAAACG-3’ (Life Technologies) were used with Roche Universal Probe Library FAM-labeled probe #58 (cat.no. 04688554001). Cpsf2-specific primers with VIC-labeled probes (Applied Biosystems; Mm00489754_m1) were used as internal control. Quantitative PCR (qPCR) was carried out using FastStart TaqMan Probe Master Mix (Roche) on software supplied with a 7500 Real-Time PCR SDS System (Applied Biosystems). PCR cycling conditions were as follows: 50°C for 2 min, 95°C for 10 min, 40 cycles at 95°C for 15 s and 60°C for 1min.
Giorgetti E., Yu Z., Chua J.P., Shimamura R., Zhao L., Zhu F., Venneti S., Pennuto M., Guan Y., Hung G, & Lieberman A.P. (2016). Rescue of metabolic alterations in AR113Q skeletal muscle by peripheral androgen receptor gene silencing. Cell reports, 17(1), 125-136.
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6

Quantifying Mitochondrial and Genomic DNA

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Genomic and mitochondrial DNA was isolated from LA/BC using the QIAamp® DNA Micro Kit (Qiagen). D-loop forward primer 5’- GACCAACATAACTGTGGTGTCA-3’ and reverse primer 5’- ATTCTTCTCCGTAGGTGCGTCTAG-3’ were designed to quantify mitochondrial DNA; Beta-2 microglobulin (B2m) forward primer 5’- TGTCAGATATGTCCTTCAGCAAGG-3’ and reverse primer 5’- TGCTTAACTCTGCAGGCGTATG-3’ were designed to quantify genomic DNA as an internal control. qPCR was carried out using SYBR Select Master mix (Life Technologies) on software supplied with a 7500 Real-Time PCR SDS System (Applied Biosystems). PCR cycling conditions were as follows: 50°C for 2 min, 95°C fo r 10 min, 40 cycles at 95°C for 15 s and 60°C for 1min. Data were expressed as Ct values and used for the relative quantification of targets with the ΔΔCt calculation to give N-fold differences. Data were transformed through the equation 2−ΔΔCt.
Giorgetti E., Yu Z., Chua J.P., Shimamura R., Zhao L., Zhu F., Venneti S., Pennuto M., Guan Y., Hung G, & Lieberman A.P. (2016). Rescue of metabolic alterations in AR113Q skeletal muscle by peripheral androgen receptor gene silencing. Cell reports, 17(1), 125-136.
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