FIELD: chemistry.
SUBSTANCE: invention relates to methods of primer-dependent propagation of nucleic acids in immobilised media, particularly to methods of detecting products of such propagation. The invention discloses an improved method of identifying changes in a nucleotide sequence and determining the number of molecules of nucleic acids as separate molecules and contained in cells or in tissue fragments. The invention enables detection of molecular colonies of nucleic acids through fluorescence of at least growth of colonies in an immobilised medium (in real time) without an additional hybridisation step and without unsealing the chamber containing the medium. If the colony is formed by more than one target molecule, the invention enables to determine the number of target molecules from which the given colony originates. This in turn enables determination of spatial location and local concentration of RNA or DNA molecules of the given type in cells or in tissue fragments contained in the gel at the polymerisation step.
EFFECT: invention can be widely used as a diagnosis method and scientific research instrument.
55 cl, 11 dwg, 12 ex
Title | Year | Author | Number |
---|---|---|---|
DETECTION OF MOLECULAR COLONIES BY MEANS OF HYBRIDISATION ON MEMBRANES WITH FLUORESCENT PROBES | 2006 |
|
RU2338787C2 |
METHOD OF NUCLEIC ACID CLONING | 1993 |
|
RU2114175C1 |
METHOD OF NUCLEIC ACID COPYING, METHOD OF THEIR EXPRESSION AND A MEDIUM FOR THEIR REALIZATION | 1992 |
|
RU2048522C1 |
METHOD OF IDENTIFICATION OF NUCLEIC ACID | 1993 |
|
RU2114915C1 |
METHOD OF INCREASING SPECIFICITY OF REVERSE TRANSCRIPTION | 2008 |
|
RU2459872C2 |
METHOD FOR PRESERVING BIOLOGICAL SAMPLE PROVIDING SAFETY OF NUCLEIC ACIDS | 2005 |
|
RU2322058C2 |
NUCLEIC ACID PROBE | 2014 |
|
RU2668154C2 |
METHOD FOR HOMOGENOUS DETECTION OF ONE-STRAND AMPLIFICATION PRODUCT | 2005 |
|
RU2460804C2 |
OLIGONUCLEOTIDES, PRIMERS AND PROBES DESIGNATED FOR REALIZATION OF DETECTION METHOD AND/OR QUANTITATIVE DETERMINATION OF ADENOVIRUS NUCLEIC ACIDS WITH USING REAGENTS SET, METHOD FOR DIAGNOSIS AND METHOD FOR SELECTION | 2001 |
|
RU2272845C2 |
SET OF OLIGODEOXYRIBONUCLETIDE PRIMERS AND FLUORESCENT-LABELED PROBE FOR IDENTIFICATION OF RNA OF HUMAN CORONAVIRUSES SARS AND 2019-NCOV BY RT-PCR METHOD WITH HYBRIDIZATION-FLUORESCENT DETECTION IN REAL TIME | 2020 |
|
RU2733665C1 |
Authors
Dates
2010-07-20—Published
2006-03-24—Filed