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A further explanation of how DNA binds to silica is based on the action of guanidinium chloride (GuHCl), which acts as a chaotrope. [3] A chaotrope denatures biomolecules by disrupting the shell of hydration around them.
Most solutions also have an antioxidant, as oxidized phenol damages the nucleic acids. For RNA purification, the pH is kept at around 4, which retains RNA in the aqueous phase preferentially. For DNA purification, the pH is usually near 7, at which point all nucleic acids are found in the aqueous phase.
Nucleic acids present in the washed (and preferably dried) silica-nucleic acid complexes is eluted into chosen elution buffer such as TE buffer, aqua bidest, and so on. The selection of the elution buffer is co-determined by the contemplated use of the isolated nucleic acid. In this way, pure nucleic acids are isolated from the starting material.
GelGreen is an intercalating nucleic acid stain used in molecular genetics for agarose gel DNA electrophoresis. GelGreen consists of two acridine orange subunits that are bridged by a linear oxygenated spacer.
He was the first scientist to isolate nucleic acid in 1869. Miescher also identified protamine and made several other discoveries. Miescher had isolated various phosphate-rich chemicals, which he called nuclein (now nucleic acids ), from the nuclei of white blood cells in Felix Hoppe-Seyler 's laboratory at the University of Tübingen , Germany ...
"Most agarose gels are made with between 0.7% (good separation or resolution of large 5–10kb DNA fragments) and 2% (good resolution for small 0.2–1kb fragments) agarose dissolved in electrophoresis buffer. Up to 3% can be used for separating very tiny fragments but a vertical polyacrylamide gel is more appropriate in this case.
A typical molecular beacon structure can be divided in 4 parts: 1) loop, an 18–30 base pair region of the molecular beacon that is complementary to the target sequence; 2) stem formed by the attachment to both termini of the loop of two short (5 to 7 nucleotide residues) oligonucleotides that are complementary to each other; 3) 5' fluorophore ...
Agarose gel is a three-dimensional matrix formed of helical agarose molecules in supercoiled bundles that are aggregated into three-dimensional structures with channels and pores through which biomolecules can pass. [3] The 3-D structure is held together with hydrogen bonds and can therefore be disrupted by heating back to a liquid state.