Liver tissue (1mg) was homogenized and RNA was isolated using Trizol (ThermoFisher, # 15596026). About 2.5μg/500 ng of total RNA was used to synthesize cDNA for mRNA/miRNA using Revertaid Reverse Transcriptase (ThermoFisher, #MAN0012885)/miSCRIPT PCR starter kit (Qiagen, #218193) respectively. Expression of immune related genes was assessed in ABI QuantStudio 7 real-time PCR machine using SYBR green (Roche). Only upregulated genes in each sample were further considered for genotyping. Primers used for quantification of mRNAs and miRNAs are presented in Supplementary Table 2A
Quantstudio 7
Manufactured by Roche
The QuantStudio 7 is a real-time PCR system designed for quantitative gene expression analysis. It features a high-resolution, touchscreen interface and supports a range of sample formats and chemistry types. The instrument is capable of performing complex thermal cycling protocols and offers data analysis capabilities.
Automatically generated - may contain errors
Lab products found in correlation
Sybr green, by Roche (2 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Revertaid reverse transcriptase, by Thermo Fisher Scientific (1 mentions)
Miscript pcr starter kit, by Qiagen (1 mentions)
Minibest universal rna extraction kit, by Takara Bio (1 mentions)
Primescript rt master mix kit, by Takara Bio (1 mentions)
2 protocols using quantstudio 7
1
Immune Gene Expression Analysis in Liver
2
RNA Extraction and qRT-PCR Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were harvested and total RNA extracted at the indicated timepoints using Takara Minibest Universal RNA extraction kit (Catalog number: 9767), then transformed to cDNA using Takara Primescript RT Master Mix kit (Takara, RR036A). The mRNA levels were then measured on an ABI (QuantStudio 7) RT-PCR system using SYBR-Green (Roche) and following manufacturer instructions. Primer sequences are shown on Table 1 .
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!