Measuring Gene Expression by RT-qPCR

Total RNA was extracted using TRIzol reagent (Invitrogen) and quantified by Nanodrop (Thermo Scientific). Real-Time PCR (RT-qPCR) was performed as previously described (51 (link)) using the SensiFAST SYBR NO-ROX One-Step (BIOLINE) following the manufacturer's instructions. Briefly, RT-qPCR reactions were performed on a Rotor-Gene Q 2plex HRM Platform (Qiagen, 9001560) and relative gene expression levels were determined using calculated concentration values, normalized to β-actin and Gapdh as reference genes. Primers used were reported in Supplementary Table S4.

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.

Maldotti M., Lauria A., Anselmi F., Molineris I., Tamburrini A., Meng G., Polignano I.L., Scrivano M.G., Campestre F., Simon L.M., Rapelli S., Morandi E., Incarnato D, & Oliviero S. (2022). The acetyltransferase p300 is recruited in trans to multiple enhancer sites by lncSmad7. Nucleic Acids Research, 50(5), 2587-2602.

Publication 2022

TRIzol reagent Nanodrop SensiFAST SYBR NO-ROX One-Step Rotor-Gene Q 2plex HRM Platform

Actin Gapdh Gene expression Genes Primers Q gene Real time pcr Trizol

Corresponding Organization : Italian institute for Genomic Medicine

Other organizations : University of Groningen

Top 5 similar protocols

Variable analysis

independent variables

RNA extraction method (TRIzol reagent)
RT-qPCR assay (SensiFAST SYBR NO-ROX One-Step)

dependent variables

Relative gene expression levels

control variables

β-actin and Gapdh as reference genes

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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.

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?

Revolutionizing how scientists
search and build protocols!