RNA pull-down

RNA pull-down

Introduction:

RNA molecules can interact extensively with other biological molecules, such as protein, DNA and RNA, and thus exist in cells and organisms in the form of macromolecular complexes. Among them, RNA-protein complexes are the existence and function of RNA One of the important forms of. RNA pull-down is a classic method for screening and identifying RNA molecules, especially the interacting proteins of lncRNA molecules, and is recognized by the majority of researchers and international authoritative academic journals. Today FORREGENE will share with you the experimental principle and technical process of RNA pull-down, maybe you can use it

1. The experimental principle of RNA pull down technology:

In vitro transcription and synthesis of biotin-labeled target lncRNA or lncRNA fragments (including RNA fragments used as positive and negative controls), and incubate with cell extracts; use avidin-anchored magnetic beads to collect RNA-protein or RNA-RNA complexes, After thorough washing, the complex is eluted. If detecting the protein target molecule that interacts with lncRNA, perform protein electrophoresis on the RNA-pull down product, detect the target protein by silver staining, and cut the gel for mass spectrometry; also directly perform western blot verification on the pull-down product; if detected For RNA target molecules that interact with lncRNA, RNA-pull down products are subjected to RNA extraction and specific qRT-PCR detection is performed.

2. RNA pull down technology experiment process:

• Technical service for RNA pull down of FOREGENE:

Cell culture

Preparation of cell total/nuclear/plasma protein extract

RNA pull down experiment

SDS-PAGE electrophoresis, silver stain

Mass spectrometry and WB experiments

Experimental report

• Experimental case

The following figure shows the results of FJ Bio-Bio's RNA pull down and SDS-PAGE electrophoresis detection, in which the red arrow marks the difference band:

The research group of Professor Song Xu of Sichuan University has accumulated 15 years of long-term accumulation in the field of lncRNA research. Foregene Bio and Professor Song Xu's research group cooperated to launch the RNA pull down technology of lncRNA, which will give you a satisfactory result for your experiment.