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karthik3k
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Does anybody have a good protocol for DNA - bead attachment ?
The Bead is streptavidin coated and the DNA has biotin and DIG on its ends.
The Bead is streptavidin coated and the DNA has biotin and DIG on its ends.
1% Blocking reagent (w/w) in
Maleic acid buffer (100 mM Maleic acid, 150 mM
NaCl, pH 7.5).
The purpose of this protocol is to attach DNA molecules to beads through the use of Streptavidin and Biotin/DIG. This allows for the isolation and purification of specific DNA sequences for further analysis.
Streptavidin is a protein that binds to biotin with high affinity. In this protocol, Streptavidin is used to bind to DNA molecules that have been biotinylated, allowing for the attachment of the DNA to the beads.
Biotinylation of DNA is important because it allows for the specific attachment of Streptavidin to the DNA molecules. This ensures that only the desired DNA sequences are isolated and purified.
DIG (Digoxigenin) is a small molecule that can be covalently attached to DNA. In this protocol, DIG is used in place of biotin to label the DNA molecules. This allows for flexibility in the attachment process and can be used in combination with biotinylated DNA.
This protocol has many applications in molecular biology, including DNA sequencing, DNA-protein interaction studies, and isolation of specific DNA sequences for cloning or further analysis. It can also be used in diagnostic tests for genetic disorders and forensic analysis.