Total RNA (500 ng) was DNAase treated, and complementary DNA was generated using the iScript complementary DNA synthesis kit (Bio-Rad Laboratories, Hercules, CA). qPCR was performed with the Applied Biosystems Incorporated 7900HT Fast Real-Time System (Life Technologies, Grand Island, NY) and LightCycler FastStart DNA Master SYBR Green as a ready-to-use reaction mix with ROX (Roche, Basel, Switzerland). Relative expression was calculated using a standard curve method, as described.39 (link),40 Results were normalized to GAPDH and fold induction compared with controls. cDNAs were amplified using TaqMan Gene Expression Assay (Hs00226845_m1) and primers (Integrated DNA Technologies, Coralville, IA) (Supplementary Table 1 ).41 (link)
Protocol detail
Quantitative Real-Time PCR Assay Protocol
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Corresponding Organization :
Other organizations : Cincinnati Children's Hospital Medical Center
Top 5 similar protocols
Variable analysis
independent variables
- None explicitly mentioned
- Relative expression (calculated using a standard curve method)
- Total RNA (500 ng)
- GAPDH (used for normalization)
- Positive control: Not specified
- Negative control: Not specified
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
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!