RNA was isolated from lung tissues using RNA isolation kit (Qiagen, Germany). After reverse transcription with Verso cDNA synthesis kit (Applied Biosystems, USA), real-time-PCR was performed on ABI 7500 system (Applied Biosystems, USA) using DyNAmoColourFlash SYBR Green qPCR kit (Thermo Fisher Scientific, USA) followed by the addition of forward and reverse primers (Integrated DNA Technologies, USA). After amplification, a melting-curve analysis was performed to verify the specificity of the reaction. The 18S rRNA gene was used as an internal control and results were determined by 2−ΔΔCt. Relative mRNA levels were expressed as the fold change over the normal control. The primer sequences were described in supplementary data (Supplementary Table S1 ).
Dynamocolourflash sybr green qpcr kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The DyNAmoColourFlash SYBR Green qPCR kit is a reagent kit designed for quantitative real-time PCR (qPCR) analysis. The kit contains SYBR Green I dye, which binds to double-stranded DNA, enabling the detection and quantification of target DNA sequences during the PCR process.
Automatically generated - may contain errors
Lab products found in correlation
Verso cdna synthesis kit, by Thermo Fisher Scientific (1 mentions)
Rna isolation kit, by Qiagen (1 mentions)
Forward and reverse primer, by Integrated DNA Technologies (1 mentions)
Abi 7500 system, by Thermo Fisher Scientific (1 mentions)
Ribozol rna extraction reagent, by Avantor (1 mentions)
Revertaid reverse transcriptase, by Thermo Fisher Scientific (1 mentions)
Thermo scientific nanodrop 2000 spectrophotometer, by Thermo Fisher Scientific (1 mentions)
Cfx96 touch real time pcr detection system, by Bio-Rad (1 mentions)
2 protocols using dynamocolourflash sybr green qpcr kit
1
RNA Extraction and Real-Time PCR for Gene Expression
2
Quantitative Gene Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total cellular RNA was extracted using Ribozol™ RNA extraction reagent (Amresco, Cleveland, OH, USA). Total RNA was quantified using Thermo Scientific™ NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). Purity of RNA was estimated by measuring the absorbance ratios at 260/280 and 260/230. Integrity of RNA was estimated using gel electrophoresis and visualizing the ribosomal RNA fractions. cDNA was synthesized using RevertAid reverse transcriptase (Thermo Fisher Scientific). qPCR was carried out using DyNAmo ColourFlash SYBR Green qPCR kit (Thermo Fisher Scientific) and specific primers in a Bio-Rad CFX96 Touch™ Real-Time PCR Detection System (Bio-Rad Laboratories Inc., Hercules, CA, USA). actb, hprt1 and polr2a were used as reference genes for qPCR. The three aforementioned genes have been identified as the most stable endogenous reference genes in the adipose tissue.16 (link) Expression levels of the target genes (abca1, slc2a4 and adipoq) were normalized to the geometric mean of the expression levels of the three reference genes.
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!