Non-radioactive marking technology for nucleic acid probes.

1. Photometric biotin markers
nucleic acids there are two types of cursor biotin
reagements
currently in use: photodynsin (acetic acid) and osteolipidin biotin. They are made up of a photosensitive group, a connecting arm, and a biotin-based group. The photo-sensitive group of photodynsin is -N3, which binds to the base in the nucleic acid under the action of light.
the photosensitive group osteolipids in osteolipid biotin can react with single-stranded or double-stranded nucleic acids under light (320 to 400 m), mainly on T, C also has a certain degree of reaction.
the connecting arm of photosensitive biotin contains 6 to 12 carbon atoms to reduce the spatial bit resistance of probe hybridization. Photosensitive biotin-labeled nucleic acids, simple method, sensitivity can also reach the pg level, can be used for
genetic
detection. Recently, a new photosensitive
dna
biotin reagent, biotin-polyethyl glycol-when-regenerative (BPA).
the DNA binding part of BPA is an active sugar-scented legume derivative that binds to the DNA base co-price bond at long-wave UVs. BPA reactants bind to DNA more specific than photosensitive biotin, which does not react with nucleic acids in visible light, a feature that allows BPA to label only nucleic acids in crude cell lysates, not proteins, polysaccharides, and other cellular molecules.2. Enzyme-based biotin-labeled nucleic acids
in place of the corresponding 32P-labeled dna by biotinized dna nucleoside triphosphate (Bio-11-dUTP, Bio-7-dATP, Bio-11-dCTP, etc.).
bio-dUTP instead of dTTP, Bio-dATP instead of dATP, Bio-dCTP instead of dCTP. Bio -11-dUTP 11 refers to the length of the carbon chain connecting the arm between the biotin group
and the deoxygenated
nucleotide. Common methods of enzymatic biotin marker DNA are notch translation and random primer extension.3. The biotin end markers of oligonucleotides
chemical markers of 5'-phosphoric acid and 3'-OH of enzymatic markers. The former connects 5'-phosphate of oligonucleotides to acetylene, and then uses amberamide biotin to connect biotin groups to the phosphate base. The latter is to add Bio-11-dUTP to its 3'-OH end (take off a pyrophosphate) with an end transfer.4. Enzyme-labeled DNA
markers are spicy root peroxidase (HRP) or alkaline
phosphatase
(AP). By connecting benzene (PBQ) to polyethylene subamine (PEI), the reagent binds to denatured DNA under the action of dialdehyde, connecting HRP to DNA to form an HRP-labeled DNA probe.5. Enzyme-labeled oligonucleotides
including nucleotide 5' end-labeled HLP and internally labeled AP. The former is to produce a -HS reaction group in HRP, which is eventually added to the 5' end of oligonucleotide synthesis, with a C6-HS base, and an active HRP reaction produces 5'-HRP oligonucleotides.
The latter is a 5' urinary glycoside 3' subphosphatidide synthesized in the oligonucleotide chain with a connecting arm and CF3 group in the process of complete oligonucleotide, after synthesis of this active oligonucleotide and AP reaction is ap-labeled oligonucleotide.6. DNA semi
antin
mark
principle is the same as Bio-11-dTUP, but the use of hairy jaundox instead of biotin to form Dig-11-dUTP, enzymatically mixed with DNA molecules. The semi-antigen molecule
Digoxigenin
marked on DNA was detected with anti-hairy jaundial antibodies .