Quantitative Analysis of Adipoq Expression

RNA was extracted from tissue using RNA STAT60 reagent (Tel-Test, Inc.) according to the manufacturer’s instructions. Synthesis of cDNA was done using TaqMan reverse transcription reagents (Thermo Fisher Scientific) using 1 μg of RNA template per reaction, as per manufacturer’s instructions. Transcript expression was then analyzed by quantitative PCR (qPCR) in 10 μL duplicate reactions using qPCRBIO SyGreen Mix (part number PB20.11; PCR Biosystems, UK) and 1–4 μL of cDNA template. Reactions were loaded into 384-well qPCR plates (part number 72.1985.202; Sarstedt, UK) and run on a Light Cycler 480 (Roche). Transcript expression was calculated based on a cDNA titration loaded on each plate and was presented relative to expression of the housekeeping gene Ppia. Primers for Adipoq and Ppia were described and validated previously (3 (link)).

Free full text: Click here

Sulston R.J., Learman B.S., Zhang B., Scheller E.L., Parlee S.D., Simon B.R., Mori H., Bree A.J., Wallace R.J., Krishnan V., MacDougald O.A, & Cawthorn W.P. (2016). Increased Circulating Adiponectin in Response to Thiazolidinediones: Investigating the Role of Bone Marrow Adipose Tissue. Frontiers in Endocrinology, 7, 128.

Publication 2016

RNA STAT60 reagent TaqMan reverse transcription reagents qPCRBIO SyGreen Mix Light Cycler 480

Cdna Housekeeping gene Light Primers Reverse transcription Synthesis Tissue Titration

Corresponding Organization : Eli Lilly (United States)

Top 5 similar protocols

Variable analysis

independent variables

RNA extraction method using RNA STAT60 reagent

dependent variables

Transcript expression analyzed by quantitative PCR (qPCR)

control variables

Housekeeping gene Ppia used as reference for relative transcript expression

controls

CDNA titration loaded on each plate as a positive control

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!