RT-qPCR was conducted using TaqMan® MicroRNA RT Kit (Applied Biosystems; Thermo Fisher Scientific, Inc.). miR-24 was used as an internal control [stem-loop RT primer sequence: 5'-GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACCTTCTG-3', forward primer: 5'-GCGGCGGTGGCTCAGTACAGC-3' for miR-24, and (universal) reverse primer: 5'-GTGCAGGGTCCGAGGT-3'] during the quantification using specific stem-loop primers for miR-15b (stem-loop RT primer sequence: 5'-GTCGTATCCAGTGCGTGTCGTGGAGTCGGCAATTGCACTGGATACGACTGTAAACC-3'; forward primer: 5'-ATCCAGTGCGTGTCGTG-3', and reverse primer: 5'-TGCTTAGCAGCACATCATG-3') to normalize it as follows: ∆Ct=∆CtmiR-∆CtmiR-24(24 (link)). The analysis of the results was performed using Sequence Detection Software version 2.3 (Applied Biosystems; Thermo Fisher Scientific, Inc.). The difference in the miR-15b relative expression levels between samples was calculated using the 2-ΔΔCq method (25 (link)).
Protocol detail
Quantifying miR-15b Expression via RT-qPCR
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.
Microrna
Primer
Stem
Stem loop sequence
Corresponding Organization :
Other organizations : Assiut University, Qassim University
Top 5 similar protocols
Variable analysis
independent variables
- MiR-15b
- MiR-15b relative expression levels
- MiR-24 (as an internal control)
- Positive control: None specified
- Negative control: None 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!