Adrenal and kidney from 8-week-old male TG mice containing either cyp11B2 haplotype-I or haplotype-II were harvested following CO2 asphyxiation and stored in All-protect tissue reagent (Qiagen). The Total RNA was isolated using RNeasy Plus minikit (Qiagen). Quantitative real-time RT-PCR (Q-RT-PCR) was next performed using power SYBR Green master mix (Cat# 4367659, Life technologies) and an ABI 7500 Fast Real-Time PCR System thermocycler (Applied Biosystems). Human (PPH01239F) and mouse (PPM57638A) CYP11B2 specific primers, and mouse GAPDH (PPM02946E) primers were purchased from SuperArray Bioscience Corporation (MD). Following a 95°C incubation for 10 minutes, 40 cycles of PCR (95°C/15s; 60°C/1m), were then performed on an ABI 7500 Fast Real-Time PCR System with 3 µl of cDNA, 50 nM PCR primers and 12.5ul SYBR Green PCR Master Mix in 25 µl reactions. Threshold cycles (CT) for three replicate reactions were determined and relative transcript abundance calculated following normalization with mouse GAPDH.
Abi 7500 fast real time pcr system thermocycler
Manufactured by Thermo Fisher Scientific
Sourced in United States
The ABI 7500 Fast Real-Time PCR System is a thermocycler designed for real-time polymerase chain reaction (PCR) analysis. It is capable of performing fast, precise, and sensitive DNA quantification and gene expression analysis.
Automatically generated - may contain errors
Lab products found in correlation
Sybr green master mix, by Thermo Fisher Scientific (1 mentions)
Rneasy plus mini kit, by Qiagen (1 mentions)
Allprotect tissue reagent, by Qiagen (1 mentions)
Nanodrop 2000, by Thermo Fisher Scientific (1 mentions)
Taqman assay, by Thermo Fisher Scientific (1 mentions)
Qubit 3.0 fluorimeter, by Thermo Fisher Scientific (1 mentions)
Reliaprep blood gdna miniprep system, by Promega (1 mentions)
Qubit dsdna br assay kit, by Thermo Fisher Scientific (1 mentions)
2 protocols using abi 7500 fast real time pcr system thermocycler
1
Quantitative CYP11B2 Expression in Mice
2
Genomic DNA Extraction and HFE Genotyping
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Genomic DNA (gDNA) was extracted from peripheral blood using the ReliaPrep Blood gDNA Miniprep System (Promega, Wisconsin, WI, USA) according to the manufacturer’s instructions. The quality of the DNA purification was verified by measuring the 260/280 (samples were ≥1.8 and ≤1.9) and 260/230 ratios (all samples > 2.0) (Nanodrop 2000, Invitrogen, Carlsbad, CA, USA). gDNA was quantified by the Qubit dsDNA BR Assay Kit (Invitrogen, California, CA, USA) in a Qubit 3.0 fluorimeter (Invitrogen, Carlsbad, CA, USA). All samples were stored at −20 °C until analyses.
HFE polymorphisms (rs1799945, C → G; and 1800562, G → A) were genotyped by TaqMan assays (Applied Biosystems, Carlsbad, CA, USA) according to the manufacturer’s instructions using an ABI 7500 Fast Real-Time PCR System thermocycler (Applied Biosystems, Foster City, CA, USA).
HFE polymorphisms (rs1799945, C → G; and 1800562, G → A) were genotyped by TaqMan assays (Applied Biosystems, Carlsbad, CA, USA) according to the manufacturer’s instructions using an ABI 7500 Fast Real-Time PCR System thermocycler (Applied Biosystems, Foster City, CA, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!