61 results about "Single copy gene" patented technology

Single copy sequences suitable for use as DNA probes can be defined by computational analysis of genomic sequences. The present invention provides an ab initio method for identification of single copy sequences for use as probes which obviates the need to compare genomic sequences with existing catalogs of repetitive sequences. By dividing a target reference sequence into a series of shorter contiguous sequence windows and comparing these sequences with the reference genome sequence, one can identify single copy sequences in a genome. Probes can then be designed and produced from these single copy intervals.

The invention provides a method of using digital PCR (polymerase chain reaction) to identify single-copy gene mutation genotypes, comprising: designing a digital PCR amplification primer and a probe for a sample's single-copy gene mutation site to be detected, designing a digital PCR amplification primer and a probe for the sample's single-copy housekeeping gene, and subjecting the mutation site and the housekeeping gene to dual-channel double digital PCR; determining whether the sample to be detected has the single-copy gene mutation site according to presence or absence of digital PCR amplification of the single-copy gene mutation site; if yes, identifying the genotype of the single-copy gene to be detected by comparing the copy number of the single-copy gene to be detected to the copy number of the single-copy housekeeping gene. The method has the advantages of good accuracy, good convenience, high speed, low cost and the like and is suitable for genetic disease screening and prevention and genetic diagnosis.

Nucleic acid (e.g., DNA) hybridization probes are described which comprise a labeled, single copy nucleic acid which hybridizes to a deduced single copy sequence interval in target nucleic acid of known sequence. The probes, which are essentially free of repetitive sequences, can be used in hybridization analyses without adding repetitive sequence-blocking nucleic acids. This allows rapid and accurate detection of chromosomal abnormalities. The probes are preferably designed by first determining the sequence of at least one single copy interval in a target nucleic acid sequence, and developing corresponding hybridization probes which hybridize to at least a part of the deduced single copy sequence. In practice, the sequences of the target and of known genomic repetitive sequence representatives are compared in order to deduce locations of the single copy sequence intervals. The single copy probes can be developed by any variety of methods, such as PCR amplification, restriction or exonuclease digestion of purified genomic fragments, or direct synthesis of DNA sequences. This is followed by labeling of the probes and hybridization to a target sequence.

The invention provides a simple and new method of using a common PCR cycler and a specific single copy gene as interior labels to realize the identification of copy numbers of transgenic plants. The invention is characterized by extracting the total DNA of young leave samples to be detected, adjusting the concentration of the DNA of all the samples to be consistent, designing specific primers of target genes and interior label genes, further determining the best PCR cycle number on the basis of PCR molecular detection, then making parallel PCR and agarose electrophoresis of the target genes and the interior label genes under the same cycle number and finally determining the copy numbers of the target genes according to the ratio of optical density values of the target genes and the interior genes in an electrophoresis image. The invention has the advantage that the invention is beneficial to screening the transgenic plants with stable expression in transgenic plant breeding; the invention has the advantages of simple operation, short time consumption, low cost, safety and accuracy; and the invention combines the PCR molecular detection of the transgenic plants and causes a laboratory for performing the PCR molecular detection of the transgenic plants to realize batch detection without adding experimental equipment and increasing the difficulty.

The invention discloses a pork component identifying molecular marker and application thereof. According to a SYBR Green I dye based real-time fluorescence quantitative PCR technology, a high-specificity and high-sensitivity pork component detection method is created by taking a pig single-copy gene in a cell nucleus as the molecular marker. By adoption of the method, pigs can be accurately distinguished from other 13 animals and 5 plants, and by practical detection of 6 meat products on the market, whether porcine-derived components are contained or not can be accurately judged. The porcine specific real-time fluorescence quantitative PCR detection method provides technical references for qualitative and quantitative detection of porcine-derived components in mixed meat products.

Nucleic acid (e.g., DNA) hybridization probes are described which comprise a labeled, single copy nucleic acid which hybridizes to a deduced single copy sequence interval in target nucleic acid of known sequence. The probes, which are essentially free of repetitive sequences, can be used in hybridization analyses without adding repetitive sequence-blocking nucleic acids. This allows rapid and accurate detection of chromosomal abnormalities. The probes are preferably designed by first determining the sequence of at least one single copy interval in a target nucleic acid sequence, and developing corresponding hybridization probes which hybridize to at least a part of the deduced single copy sequence. In practice, the sequences of the target and of known genomic repetitive sequence representatives are compared in order to deduce locations of the single copy sequence intervals. The single copy probes can be developed by any variety of methods, such as PCR amplification, restriction or exonuclease digestion of purified genomic fragments, or direct synthesis of DNA sequences. This is followed by labeling of the probes and hybridization to a target sequence.

The invention discloses a method for identifying the age of ginseng by utilizing the length of a ginseng telomere. The method comprises the following steps: (A) establishing a mathematical model: (a1) selecting a plurality of samples from the positions 1-2 cm below the rhizomes of the ginseng of known age and extracting genomic DNA from each sample; (a2) taking each genomic DNA as a template, conducting qPCR on a primer pair distinguished from the ginseng telomere to obtain a Ct value and marking the Ct value as T, and conducting qPCR on a primer pair distinguished from a single copy gene of the ginseng to obtain another Ct value and marking the Ct value as S; (a3) calculating the T / S ratio for each ginseng; (a4) taking the ages of the ginseng as the abscissa values and taking all the calculated T / S ratios as the ordinate values to draw a standard linear curve and obtain a curve equation; (B) adopting the mathematical model to identify the age of ginseng to be measured: obtaining the T / S ratio for the ginseng to be measured according to the operations from step (a1) to step (a3), and substituting the T / S ratio into the mathematical model to obtain the age of the ginseng to be measured. The method has the advantages that the identified age of ginseng is highly identical with the actual age of the ginseng, the cost is low, and the flux is high. Compared with the traditional Southern blot technology adopted for measuring the length of a ginseng telomere, the method provided by the invention has less demand for DNA and a shorter test period.

The invention relates to a quantitative detection method for donkey-derived and swine-derived components in a donkey-hide gelatin liquid semi-finished product or finished product, a composition and a kit, which belongs to the technical field of molecular biology. The method comprises: according to a donkey and swine house-keeping gene single-copy gene, designing a universal donkey and swine primer as well as a donkey and swine specificity probe, extracting a sample DNA to be tested, quantitatively detecting the donkey-derived or swine-derived components in the donkey-hide gelatin liquid semi-finished product or finished product by adopting a TaqMan qPCR method, calculating a copy number of donkey-derived and swine-derived karyogene in the unit mass of the donkey-hide gelatin liquid according to a standard curve and a Ct value of a sample to be tested, calculating a ratio of the donkey-derived components and a ratio of the swine-derived components in donkey-hide gelatin, and quantifying the donkey-derived or swine-derived components in the gelatin sample. The method, the composition and the kit are good in detection specificity, high in sensitivity, not only capable of qualitatively detecting the donkey-derived and swine-derived components in the donkey-hide gelatin, but also capable of realizing the quantitative analysis, and capable of providing technical popularization and application for the quality control of donkey-hide gelatin enterprises.

The invention discloses a method for quantitatively detecting nuclear genome copy number and human mitochondrial genome copy number by fluorescence. The method includes respectively selecting a singlecopy gene in human nuclear genome DNA and mitochondrial genome, designing primers and probes, transferring sequence fragments into PMD-18T vectors, extracting plasmids after conversion to obtain thecopy number and taking the copy number as a reference standard; and in actual detection, taking the Cq value of the reference standard to make a standard curve to compare and obtain the copy number ofnuclear genome DNA and mitochondrial genome DNA in a sample to be detected. The method provides a method for quantitatively analyzing the copy number of the mitochondrial genome in different human tissue cells, has high specificity and simple operation, and can complete rapid detection of the copy number of the mitochondrial genome only by a fluorescence quantitative PCR instrument.

The invention discloses a primer pair for detecting and identifying Pectobacterium bacteria and a using method thereof. The primer comprises a forward primer 23SPecF and a reverse primer 23SPecR. The method for detecting and identifying the Pectobacterium bacteria comprises the following steps of: performing Polymerase Chain Reaction (PCR) on the 23SPecF and the 23SPecR by using the primer pair, wherein a 266bp nucleic acid fragment can be only amplified from a sample containing the genome DNA of the Pectobacterium bacteria. Due to that the Pectobacterium bacteria have seven copied 23SrRNA genes, the detection sensitivity of the primer pair for performing the PCR on the 23SPecF and the 23SPecR is seven times that of a single-copy gene primer, and the Pectobacterium bacteria can be detected more sensitively.

The invention discloses a composite amplification kit for simultaneously detecting 39 human Y chromosome STR loci. 39 Y chromosome loci are simultaneously amplified in a composite amplification reaction system. The composite amplification kit comprises 26 single-copy loci, 3 double-copy loci, one three-copy loci and one four-copy loci. The 39 loci are divided into five groups through a loca arrangement mode which is unique in design and a specific primer sequence, and each group is labeled with a different fluorescent label. The kit has high resolution and high individual identification amongmale samples.

The invention discloses a recombinant alginate lyase, its engineering bacteria and a method for preparing alginate oligosaccharides by hydrolysis. According to the newly isolated strain Vibrio sp.BZM‑1 from seaweed, the alginate gene-AlgM is screened; The expression vector of the gene, pHBM905BDM‑AlgM, is transformed into Pichia pastoris GS115 competent cells to obtain single-copy genetically engineered bacteria; construct, culture and verify multi-copy genetically engineered bacteria, efficiently express recombinant alginate lyase, and use this enzyme to hydrolyze alginic acid Sodium production of fucoidan oligosaccharides. GS115 / AlgM4 (CCTCC No: M2015500) with four copies of recombinant alginate lyase genetically engineered strain had a higher expression level. The activity of the recombinant alginate lyase of the present invention is as high as 31000U / mL.

Single copy sequences suitable for use as DNA probes can be defined by computational analysis of genomic sequences. The present invention provides an ab initio method for identification of single copy sequences for use as probes which obviates the need to compare genomic sequences with existing catalogs of repetitive sequences. By dividing a target reference sequence into a series of shorter contiguous sequence windows and comparing these sequences with the reference genome sequence, one can identify single copy sequences in a genome. Probes can then be designed and produced from these single copy intervals.

Nucleic acid (e.g., DNA) hybridization probes are described which comprise a labeled, single copy nucleic acid which hybridizes to a deduced single copy sequence interval in target nucleic acid of known sequence. The probes, which are essentially free of repetitive sequences, can be used in hybridization analyses without adding repetitive sequence-blocking nucleic acids. This allows rapid and accurate detection of chromosomal abnormalities. The probes are preferably designed by first determining the sequence of at least one single copy interval in a target nucleic acid sequence, and developing corresponding hybridization probes which hybridize to at least a part of the deduced single copy sequence. In practice, the sequences of the target and of known genomic repetitive sequence representatives are compared in order to deduce locations of the single copy sequence intervals. The single copy probes can be developed by any variety of methods, such as PCR amplification, restriction or exonuclease digestion of purified genomic fragments, or direct synthesis of DNA sequences. This is followed by labeling of the probes and hybridization to a target sequence.