Validating RNA-Seq Findings by qPCR

To validate the findings of the RNA-Seq assay, 20 DEGs were randomly chosen and their relative expression confirmed by qPCR analysis using the fluorescent intercalating dye SYBRGreen in the Opticon 2 detection system (MJ Research, Waltham, MA, USA). Details of the selected genes and the respective primers are listed in Table S1. Gene expression levels were normalized against the hulless barley gene HvADP (Ferdous et al., 2015 (link)) and calculated using the 2^−ΔΔCT method (Livak and Schmittgen, 2001 (link)). Three technical replicates were generated for each biological sample.

Free full text: Click here

Wei Z., Zeng X., Qin C., Wang Y., Bai L., Xu Q., Yuan H., Tang Y, & Nyima T. (2016). Comparative Transcriptome Analysis Revealed Genes Commonly Responsive to Varied Nitrate Stress in Leaves of Tibetan Hulless Barley. Frontiers in Plant Science, 7, 1067.

Publication 2016

Opticon 2

Assay Barley Biological Dye 2 Fluorescent dye Gene Gene expression Primers Rna seq

Corresponding Organization : Tibet Academy of Agricultural and Animal Husbandry Sciences

Top 5 similar protocols

Variable analysis

independent variables

RNA-Seq assay

dependent variables

Relative expression of 20 DEGs confirmed by qPCR analysis using SYBRGreen in the Opticon 2 detection system

control variables

Gene expression levels normalized against the hulless barley gene HvADP

controls

Positive control: Not specified
Negative control: Not 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.

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!