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The specificity of the hybridization-ligation assay for ligation at the 3'-end is particularly relevant because the predominant nucleases in blood are 3' to 5' exonucleases. One limitation of the method is that it requires a free 3'-end hydroxyl which may not be available when targeting moieties are attached to the 3'-end, for example.
Contamination by phenol, which is commonly used in nucleic acid purification, can significantly throw off quantification estimates. Phenol absorbs with a peak at 270 nm and a A 260/280 of 1.2. Nucleic acid preparations uncontaminated by phenol should have a A 260/280 of around 2. [2]
Boom method (aka Boom nucleic acid extraction method) is a solid phase extraction method for isolating nucleic acid from a biological sample. This method is characterized by "absorbing the nucleic acids (NA) to the silica beads".
Deoxyribonucleic acid (DNA) is a nucleic acid containing the genetic instructions used in the development and functioning of all known living organisms. The chemical DNA was discovered in 1869, but its role in genetic inheritance was not demonstrated until 1943. The DNA segments that carry this genetic information are called genes.
[7] For visualization purposes, the nucleic acid fragment is usually labelled with a radioactive, fluorescent or biotin label. Standard ethidium bromide staining is less sensitive than these methods and can lack the sensitivity to detect the nucleic acid if small amounts of nucleic acid or single-stranded nucleic acid(s) are used in these ...
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 ...
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.
Nucleic acid sequence-based amplification, commonly referred to as NASBA, is a method in molecular biology which is used to produce multiple copies of single stranded RNA. [1] NASBA is a two-step process that takes RNA and anneals specially designed primers, then utilizes an enzyme cocktail to amplify it.