RNA Isolation and qPCR Analysis

Tissues or cells were lysed in TRIzol, and RNA was isolated using an Invitrogen™ PureLink RNA Mini Kit (Cat No: 12183025, ThermoFisher) according to the manufacturer’s instructions. RNA concentration was determined by NanoOne, and an equal amount of RNA was used to prepare cDNA using LunaScript® RT SuperMix (New England BioLabs, Cat no: M3010L) according to the manufacturer’s conditions. qPCR was performed by TaqMan primer probes using Luna Universal Probe qPCR Master Mix (New England Biolabs, Cat No: M3004) and Bio-Rad i-cycler according to standard lab procedures^{9 (link),10 (link),31 (link)}. HPRT was used to normalize gene expression^{31 (link),32 (link)}. The ∆∆Ct method was used to calculate relative gene expression values in qPCR analysis.

Free full text: Click here

Batan S., Kuppuswamy S., Wood M., Reddy M., Annex B, & Ganta V. (2024). Inhibiting anti-angiogenic VEGF165b activates a miR-17-20a-Calcipressin-3 pathway that revascularizes ischemic muscle in peripheral artery disease. Communications Medicine, 4, 3.

Publication 2024

Invitrogen™ PureLink RNA Mini Kit LunaScript® RT SuperMix Luna Universal Probe qPCR Master Mix

Corresponding Organization : Augusta University

Top 5 similar protocols

Variable analysis

independent variables

Not explicitly mentioned

dependent variables

Gene expression

control variables

HPRT expression used to normalize gene expression

controls

Positive control: None mentioned
Negative control: None 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!