What is biotinylated DNA?
In biochemistry, biotinylation is the process of covalently attaching biotin to a protein, nucleic acid or other molecule. Biotinylation is rapid, specific and is unlikely to disturb the natural function of the molecule due to the small size of biotin (MW = 244.31 g/mol).
What is RNA pull down assay?
RNA pull-down assays selectively extract a protein–RNA complex from a sample. Typically, the RNA pull-down assay takes advantage of high affinity tags, such as biotin or azido-phosphine chemistry.
Can DNA be biotinylated?
Other nucleic acid biotinylation techniques The photoactivatable biotin can be incorporated randomly in the DNA fragment double-stranded DNA and single-stranded DNA or RNA. Photoactivatable biotin is simply added to the sample and irradiated with UV light.
Why is biotinylation important?
The biotin–avidin interaction is commonly exploited to detect and/or purify proteins because of the high specificity that these two molecules have for each other. Biotinylation is the process of attaching biotin to proteins and other macromolecules.
How do you identify RNA?
Alternatively, the protein may be labeled, or the RNA–protein complex may be isolated using an antibody against the protein of interest. The RNA is then detected by northern blot or through RT-PCR analysis and the proteins detected by western blotting or mass spectrometry.
What is Desthiobiotin?
Desthiobiotin is a biotin analogue that binds less tightly to biotin-binding proteins and is easily displaced by biotin. We synthesized an amine-reactive desthiobiotin derivative for labeling proteins and a desthiobiotin-agarose affinity matrix.
Why is biotin used in Elisa?
Streptavidin-biotin technology has been effectively used to improve ELISA detection due to the strong affinity between biotin and streptavidin because it has a dissociation constant (Kd) in the femtomolar range. Because biotin and streptavidin interact strongly, more analyte molecules can be captured on an ELISA plate.
How to improve the biotinylation efficiency of RNA?
Description. Optimization of biotinylation efficiency for complex RNA molecules is achieved by altering the RNA to biotinylated cytidine ratio, increasing the incubation time or by adding DMSO to the labeling reaction in order to denature the RNA structure. Following the RNA biotinylation reaction, labeled RNA is easily precipitated…
How are biotin labels used in RNA biotinylation?
Following the RNA biotinylation reaction, labeled RNA is easily precipitated to remove reaction byproducts. The biotin label enables the RNA probe to be immobilized or detected with streptavidin in order to study RNA interactions (i.e., RNA electrophoretic mobility shift assays, RNA pull-down assays) or to perform nonradioactive blotting…
What is Thermo Scientific Pierce RNA 3’end biotinylation kit?
The Thermo Scientific Pierce RNA 3′ End Biotinylation Kit is optimized for labeling the 3-prime terminal end of RNA probes to facilitate their use as probes or targets in EMSA and other methods for studying protein-RNA interactions.
How are biotinylated proteins used in molecular biology?
Due to the strong affinity between biotin and streptavidin, the purification of biotinylated proteins has been a widely used approach to identify protein-protein interactions and post-translational events such as ubiquitylation in molecular biology. Proteins can be biotinylated chemically or enzymatically.