RNA Extraction and qPCR Analysis

Total RNA was extracted from the tumor cells and tissues using TRIzol (Sigma) reagent according to a previously described protocol 29 (link). A NanoDrop 2000 (Thermo, USA) was used to quantify the concentrations of the RNA. Total RNA was then reverse transcribed to complementary DNA (cDNA) using a miScript II RT kit (TaKaRa Biotechnology Co., Ltd., Dalian, China), and qPCR was performed using TB Green® Premix Ex Taq™ (RR820A, Takara, Japan) and a Bio-Rad real-time PCR instrument.
The primer sequences are listed in Supplementary Table 1.

Free full text: Click here

Wu W., Li Q., Zhu Z., Li C., Lu P., Zhou X., Huang Y., Liu Y., Wang M, & Gong J. (2022). HTR1D functions as a key target of HOXA10-AS/miR-340-3p axis to promote the malignant outcome of pancreatic cancer via PI3K-AKT signaling pathway. International Journal of Biological Sciences, 18(9), 3777-3794.

Publication 2022

TRIzol NanoDrop 2000 miScript II RT kit TB Green® Premix Ex Taq™ real-time PCR instrument

Cdna Primer Real time pcr Tissues Trizol Tumor

Corresponding Organization : Second Affiliated Hospital of Chongqing Medical University

Other organizations : First Affiliated Hospital of Chongqing Medical University, Affiliated Hospital of Southwest Medical University, Chengdu Fifth People's Hospital

Top 5 similar protocols

Variable analysis

independent variables

Total RNA extraction using TRIzol reagent
Reverse transcription of total RNA to cDNA using miScript II RT kit
QPCR analysis using TB Green® Premix Ex Taq™ and Bio-Rad real-time PCR instrument

dependent variables

Concentrations of the extracted RNA measured using NanoDrop 2000

control variables

Previously described protocol 29 (link) for total RNA extraction

controls

No positive or negative controls were explicitly mentioned in the provided information.

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!