RT-qPCR for mRNA and miRNA Analysis

Total RNA was extracted by Trizol reagent (Invitrogen) and then was used to synthesize cDNA by Moloney murine leukemia virus reverse transcriptase (New England BioLabs) with an oligo (dT)₁₈ primer. According to the manufacturer's protocol (Applied Biosystems), the cDNAs were amplified with primers listed in the Supplementary Table S1 using an ABI StepOne Plus system with SYBR Green Master Mix for mRNA detection [43 (link)]. For detection of mature miRNAs, cDNA synthesis was carried out by MultiScribeTM Reverse Transcriptase system and then quantitative miRNA real-time PCR was performed by the ABI StepOne Plus system (Applied Biosystems) [42 (link)].

Free full text: Click here

Hsu K.W., Fang W.L., Huang K.H., Huang T.T., Lee H.C., Hsieh R.H., Chi C.W, & Yeh T.S. (2016). Notch1 pathway-mediated microRNA-151-5p promotes gastric cancer progression. Oncotarget, 7(25), 38036-38051.

Publication 2016

Trizol reagent Moloney murine leukemia virus reverse transcriptase ABI StepOne Plus system SYBR Green Master Mix

Cdna Mirna Moloney murine leukemia virus Mrna Oligo Primers Quantitative real time pcr Reverse transcriptase Sybr green Synthesis Trizol

Corresponding Organization : Institute of Molecular Biology, Academia Sinica

Other organizations : China Medical University, China Medical University Hospital, Taipei Veterans General Hospital

Top 5 similar protocols

Protocol cited in 1 other protocol

Variable analysis

independent variables

Primer design for mRNA detection
Use of SYBR Green Master Mix for mRNA detection
Primer design for miRNA detection
Use of MultiScribeTM Reverse Transcriptase system for miRNA cDNA synthesis

dependent variables

MRNA expression levels
MiRNA expression levels

control variables

Total RNA extraction by Trizol reagent
CDNA synthesis by Moloney murine leukemia virus reverse transcriptase with oligo (dT)18 primer
Use of ABI StepOne Plus system for mRNA and miRNA detection

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!