METHOD OF SUPPRESSING FORMATION OF PHOTOCROSSLINK, AND PHOTOREACTIVE NUCLEIC ACID IN WHICH AUTO-CROSSLINK FORMATION IS SUPPRESSED
申请人:JAPAN SCIENCE AND TECHNOLOGY AGENCY
公开号:US20160326207A1
公开(公告)日:2016-11-10
Provided is a means for preventing the inactivation of a photoresponsive nucleic acid probe by suppressing the formation of a photocrosslink between a photoresponsive base having a photocrosslinkable vinyl structure and a photocrosslinkable thymine (T) or uracil (U) base, by substituting with an R group (R being —CN or —CO—R
1
, where R
1
is a saturated or unsaturated straight-chain or branched cyclic or non-cyclic C1-12 hydrocarbon group) the 5 position of a pyrimidine ring of the thymine (T) or uracil (U) base which is photocrosslinkable with the photoresponsive base having a photocrosslinkable vinyl structure.
A method of detecting a target nucleic acid by sandwich hybridization using a detection probe that hybridizes with the target nucleic acid, and a capture probe immobilized on a support wherein at least one nucleic acid base in the nucleic acid molecule(s) of the detection probe and/or capture probe with a photoreactive group includes irradiating, with light, a complex formed by hybridization between the target nucleic acid and the detection probe and/or capture probe to allow formation of a covalent bond(s) between the photoreactive group(s) and a nucleic acid base(s) in the target nucleic acid.
The present invention relates to a nucleic acid linker for producing a complex of mRNA, and a protein or a peptide which is encoded by the mRNA, the linker comprising: a spacer portion at the 5′-terminal; a polynucleotide portion hybridizable with at least a part of a sequence of the mRNA; and an arm portion which has a connection portion for the protein or the peptide at the 3′-terminal, in which the spacer portion, the polynucleotide portion, and the arm portion form a single strand, and in which the polynucleotide portion contains a photoreactive base derivative.
A method for detecting a target nucleic acid comprising: forming a three-component association product by allowing the association of at least a nucleic acid molecule, a first nucleic acid probe having a first marker bound thereto, and a second nucleic acid probe having a second marker bound thereto; forming at least one covalent bond between the target nucleic acid molecule and the first nucleic acid probe and between the target nucleic acid molecule and the second nucleic acid probe; and binding the three-component association product to a solid phase carrier through the second marker; recovering the three-component association product bound to the solid phase carrier; releasing the first marker from the recovered three-component association product; and detecting the target nucleic acid molecule by detecting the free first marker.
PHOTOCOUPLING METHOD USING PROBE CONTAINING PHOTORESPONSIVE NUCLEIC ACIDS
申请人:LSI Medience Corporation
公开号:EP2878671A1
公开(公告)日:2015-06-03
Provided are: a photocoupling method that overcomes the problem of the stagnation of photocoupling with a target nucleotide using a probe containing a photo-responsive nucleotide, and that improves the photocoupling efficiency; and a photocoupling kit.
The photocoupling method is characterized by hybridizing a target site present in a nucleic acid sample with a first probe having a sequence complementary to the target site and containing a photo-responsive nucleotide, in a reaction solution, and carrying out photocoupling by photo-irradiation, wherein self-assembly caused by the photo-responsive nucleotide contained in the first probe is suppressed. The photocoupling kit is characterized by comprising a first probe having a sequence complementary to a target site present in a nucleic acid sample, and containing a photo-responsive nucleotide; and a second probe being highly complementary to the first probe.