RNA Expression Analysis by qPCR

RNA was isolated using RNeasy mini kit with on-column DNase I digestion (Qiagen). Reverse transcription followed by qPCR was performed by preparing cDNAs with Prime Script RT Master Mix (Takarabio) or QuantiTect RT-Kit (Qiagen), and qPCR with SYBR Green reagent using indicated gene primers (Supplemental Table 4) on a 7500 Fast Real-Time PCR system (Thermo Fisher Scientific). Data were analyzed using the 2^–ΔΔCt method to determine relative gene expression after normalization with internal control genes (RPLPO, HPRT1, and PPIA for human) and (Rplp0 and Ppia for mouse genes) (21 (link), 22 (link)).

Free full text: Click here

Reddy M.A., Amaram V., Das S., Tanwar V.S., Ganguly R., Wang M., Lanting L., Zhang L., Abdollahi M., Chen Z., Wu X., Devaraj S, & Natarajan R. (2021). lncRNA DRAIR is downregulated in diabetic monocytes and modulates the inflammatory phenotype via epigenetic mechanisms. JCI Insight, 6(11), e143289.

Publication 2021

RNeasy mini kit on-column DNase I digestion Prime Script RT Master Mix QuantiTect RT-Kit 7500 Fast Real-Time PCR system

Cdnas Digestion Dnase i Gene expression Genes Human Mouse Primers Reverse transcription Rplp0 Sybr green

Corresponding Organization : City of Hope

Other organizations : Central Drug Research Institute, Texas Children's Hospital, Baylor College of Medicine

Top 5 similar protocols

Variable analysis

independent variables

RNA isolation method (RNeasy mini kit with on-column DNase I digestion (Qiagen))
Reverse transcription method (Prime Script RT Master Mix (Takarabio) or QuantiTect RT-Kit (Qiagen))
QPCR method (SYBR Green reagent)

dependent variables

Relative gene expression

control variables

Internal control genes (RPLPO, HPRT1, PPIA for human; Rplp0, Ppia for mouse)

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!