IDENTIFICATION MARKERS FOR HANWOO AND CHINESE YANBIAN VARIETIES USING DNA REPLICATION NUMBER VARIATION AND IDENTIFICATION METHOD THEREOF
The present invention refers to DNA garment number identification marker to be Chinese bred by the same and method of identification of shadow area along varieties and method are disclosed. More specifically cattle can be exhibited in a shadow area along China differentially populations can analyze specific gene sequence number 1 to 6 number of transitions be variant gene marker [...] prevention and number, in a quicker and more accurate and using the same, and pretensioned subtract the ID number to identify a China cattle area along diameter method are disclosed. The varietal 2,000 years or more unique Korean cattle in Korean peninsula which compares the output signal independently on world unique to the varietal, the Korean foreign bovine (shadow area along China, Europe [...], like it is a hall star) is bred by the same varieties is differently than when the varietal evolving, the flesh of the gene are associated with value in cattle is reported that there are, according to the unique development of meat and meat amount is up. The domestic flow varieties [...] cattle meat, meat and revenue and six cow, cattle meat sold at higher consumption of cattle for pork consumer often while the tenderness revenue, the reliability (in particular adjacent the shadow area along China Korean) non-cattle cattle varietal identification a need for last year. The cattle (Korean China in particular adjacent the shadow area along) the decreasing cattle and non specific marker gene develop cattle, using PCR DNA integrity of polymorphic RAPD (random amplified polymorphic DNA), cattle and specific surface of the MC1R gene identification method using techniques through fitting, and PCR-a RFLP (restriction fragment polymorphism) retrieving an appropriate gene marker studies using a method is not disclosed. However PCR of DNA polymorphism analysis using RAPD (random amplified polymorphic DNA) pictures, wide varieties or species identification stage to various animal genetic analyses and applications, accuracy or reproducibility of the pin is difficult practical result low drainage holes. Related patent patent 10 - 0871145 to PCR using specific SNP is Korean registration on both determining method is disclosed for determining the bending machine species bovine species. The purpose of the invention to cattle area along raincoat species and the China cattle can be exhibited in a cattle and China area along the shadow area along China differentially population specific marker gene or hypermetropia [...] number number be a backhoe is desired. It is another object of the present invention area along China cattle for cattle raincoat species and the specific gene can be exhibited in a shadow area along China differentially population number be kits including cattle and China marker number UGT [...] prevention area along a backhoe. Said gene of the present invention another object is to provide a rapid number which are arranged mirror number than be prevention China area along the varietal identification method number [...] a raincoat species and allow cattle are disclosed. Another object of the present invention and the advantage of the following detailed description of the invention, claims and drawing by more specifically with each other. One aspect of the present invention, sequence number 3 gene variant markers identifying number represented by probe including bovine species be encoded ball number is desired. One aspect of the present invention, sequence number 6 probe including bovine species is indicated number identifying number encoded gene variant markers be prevention ball. In the embodiment of the invention is the one, characterized in that said it is right area along China cattle cow number identifying number encoded gene variant markers be small the varietal prevention ball. By another aspect of the present invention is, represented by sequence numbers 1 and 2 which enables the varietal composition including bovine encoded identifying number ball. By another aspect of the present invention is, represented by sequence numbers 4 and 5 which enables identifying composition including bovine species encoded ball number. The present invention is to an in the embodiment, it is right area along said China cattle cow characterized encoded identifying number ball small varietal composition. Another aspect of the present invention, said composition including ball number encoded kits. By another aspect of the present invention is, subject nucleic acid sample separation step; said isolated nucleic and IIS, marker gene sequence number for primer set or sequence number 1 and 2 identify and amplify prevention be number 4 and 5 amplified target sequence to yield for primer set for amplifying an amplification step; and said amplified gene product by positive number (Copy) confirming including bovine species identification method is number number number desired etched hole. In the embodiment of the present invention is one, it is right area along the varietal identification method is characterized in that said cattle cow China or small ball number encoded. In the embodiment of the present invention the one, said gene number number identifying step ddPCR (Droplet-a digital PCR) prevention analyzing method is characterized by using the varietal identification number encoded small ball. In the embodiment of the present invention the one, said gene number number identifying said composition prevention step, determining if the varietal identification method number be less than 1 weight fluctuations area along the rain and dew prevention is encoded small ball number. In the embodiment of the present invention the one, cattle and China marker gene be available can be distinguish area along [...] number number is desired. In the embodiment of the present invention the one, cattle and China area along the varietal available distinguish primer set is included [...] identifying composition can be a small number. In the embodiment of the present invention the one, said number can be [...] kits is the composition. In the embodiment of the present invention is one, the varietal identification method can distinguish cattle and China area along a small number can be [...]. The varietal cows using said gene marker and composition in a quicker and more accurate method is to provide a method to identify the varieties available identification number and diameter, revenue meat demand increase to accommodate China cattle by identifying number [...] differentiation applicable to livestock management elementary bounds area along prevention can be improved disclosed. Figure 1 cattle (N=7) (N=7) 2 in China marker representing the result possibility mutation on two garment number ddPCR shadow area along are disclosed. Figure 2 is representing a distribution area along cattle are found in the genome of a non-volatile memory CNVR, [...] and green source only obtain a shadow only obtain a big cattle or area along each. The size of the won CNVR indicating a density linear correlation are disclosed. Figure 3 cattle CNVR raincoat area along the size distribution in the chemical formula, the size of the bar graph includes a CNVR (<5 kb, 10 kb<, 15 kb<,<20 kb,<30 kb, and>30 kb, X axis) of the present invention genic provided CNVR CNVR estimated for an entire size distribution (A) and indicating the distribution of (B) are disclosed. The present invention more comprehensible [...] for specific terms herein defined as follows. Alternatively herein defines not, scientific terms and techniques used in the present invention terms are read by the meanings in the art generally will have a person with skill in the art are disclosed. In addition, specifically providing language translators which does not specify a, in the form of a plurality of aspect of the same term which rapidly including, in the form of a single aspect of the term is understood to including a plurality it should. Hereinafter, the present invention more detailed as follows. One aspect of the present invention, sequence number 3 or number identifying gene probe is indicated including bovine species can be [...] variant marker number is desired. Used in the present invention 'gene variant (Copy-a number variation, CNV) be desired number' lymphocytes transitions corresponding to transitions structures in such that only one genetic change, dielectric structural variant (structural variation) observed in a smallest recoat disclosed. Typically in the form of a generally 2n unlike deletion sequences (0n, 1n state), amplification (3n or more of the States) compared to the standard reference genome such as representative (Reference Genome) 1 kb or more DNA sequence repeatedly show differences in the number of pieces which primarily means, 29 kb 523 kb average size reported in extent over [Pawel Stankiewicz, James R. Lupski (2010). "Variation in the Human Genome and its Role in Disease" 61 "Annual Review of Medicine": 437455.]. One aspect of the present invention, sequence number 6 probe including bovine species is indicated number identifying number can be [...] variant marker gene be desired. In possibility mutation on chromosome number system capable of shadow area along China cattle varieties for joint and area is desired, the applicant cattle (N=10) on the China area along each individual data according to a next generation of shadow (N=10) when the noise average 10 producing bio-sequence listing (Next Generation sequencing), garment number for later possibility mutation region of garment number utilizing software CNVSeq possibility mutation found total 901. One of the, number of difference I be in shadow area along side when the number is China cattle possibility mutation marker number 2 prevention prevention (CNVR_13_11, CNVR_8_24) for ddPCR cattle (N=7) (N=7) population number utilized to predict variation on shadow area along China conducting verification experiment number is desired. In the embodiment of the invention is the one, characterized in that said it is right area along China cattle cow number can be [...] number be small the varietal identifying variant marker gene is desired. It cries cattle appearance or similar area along China, recent experience during year tens to hold the varietal and other livestock, in the case of cattle, which affects the growth and China on right area along that preferentially with a larger number of component is generated from the China cattle etc. need increased area along raincoat varieties of identification. The present invention refers to a non-volatile memory area along cattle full genome garment number comparison of sequencing data identifying number of variant (genome-a wide copy number variations) be total 901 prevention be shifting region (CNVRs) and thus, the present invention be a larger difference between them the arrears of work to find a number variation is desired. By another aspect of the present invention is, represented by sequence numbers 1 and 2 which enables identifying composition can be [...] number including bovine species. The term "primer" is used in the present invention for free 3 'end hydroxyl radicals (free 3' hydroxyl group) having radiation nascent strand sequence starting point function by big short sequences. Appropriate buffer solution at temperature and polymerization primer (i.e., DNA polymer produced or reverse transcriptase) 4 in the presence of nucleoside triphosphate is different reagents and for DNA synthesis can be disclosure. PCR amplification of the desired product through specific embodiment can be predicting whether a diagnosis of abdominal obesity. PCR conditions, the contrast based on the length of the antisense primer into a sense can be publicly known. By another aspect of the present invention is, represented by sequence numbers 4 and 5 which enables identifying composition including bovine species can be [...] number. The present invention is to an in the embodiment, it is right area along said China cattle cow characterized composition can be [...] small varietal identifying number. Another aspect of the present invention, said composition including kits can be [...] number. If kit of the present invention performs amplification PCR when applied, the kits of the present invention selectively, PCR amplification reagents required, for example, buffer, DNA polymerase (e.g., Thermus aquaticus (Taq), Thermus thermophilus (Tth), Thermus filiformis, Thermis flavus, Thermococcus literalis or Pyrococcus furiosus DNA polymerase (Pfu) obtained from thermal stability), a DNA polymerase can be cofactors and dNTPs, if applied to a immunoassay kit of the present invention, kit of the present invention selectively, secondary antibodies and comprising determination of substrate can be. Further, the present invention according to said agent component including a plurality of separate com part with distant [thu kit packaging or smaller number can be disclosed. By another aspect of the present invention is, subject nucleic acid sample separation step; said isolated nucleic and IIS, marker gene sequence number for primer set or sequence number 1 and 2 identify and amplify prevention be number 4 and 5 amplified target sequence to yield for primer set for amplifying an amplification step; and said amplified positive number gene product by a method including the varietal identification number (Copy) confirming the number prevention can be bovine [...] number. Said nucleic acid can be single or double-stranded form, RNA, DNA, or backbone modified ribonucleotides or deoxyribonucleotides - per - can be a molecule such as nucleic acid analogs, the number is not one. In the embodiment of the present invention is one, it is right to the varietal identification method characterized cattle or China area along said cow [...] number can be small. In the embodiment of the present invention the one, said gene number number identifying step ddPCR (Droplet-a digital PCR) prevention using a method analyzing small varietal identification number can be characterized [...]. Terms used in the present invention 'ddPCR (Droplet Digital PCR)' in the qPCR 2 generation PCR (quantitative PCR (real-a time PCR) the efficiency of the PCR result value according to the device for that can be caused by the database, the number of droplets after PCR amplification of cleavable [...] 20ul 1nl reaction, can be counting the target DNA. In addition Target DNA amplification of positive according to the distribution of negative charge only calculate [...] Target DNA copy by wind, and by means of which a standard curve of quantitative PCR that is cleavable with absolute quantitative PCR reaction of decoded video without droplets separated from and has higher sensitivity and accuracy of qPCR, according to PCR efficiency not affected constant results can be achieved. In the embodiment of the present invention the one, said gene number number identifying said composition prevention step, determining the varietal identification method number be less than 1 area along the rain and dew fluctuations if desired a small number can be [...]. As a result, possibility mutation markers present we shall be in shadow population number [...] CNVR_13_11 garment number area along prevention, prevention of further present in cattle population number [...] 1 has been confirmed. In addition, the biomarker is an area along the shadow population average 0 CNVR_8_24 garment number possibility mutation. 002 Prevention support number has two be transitions, in cattle population average 58. 12 [...] which shows two garment number has been confirmed. Hereinafter, the present invention in the embodiment by a detailed the on-sensors other. In the embodiment these more specifically account for the present invention is provided, in the embodiment of the present invention range and not the limited to these. In the embodiment 1. Cold rolled material and Genomic DNA extraction This cattle (HanWoo) 7 used in the cold rolled material is two, China (Chinese Yanbian cattle) 7 shadow area along two, two blood DNA total 2 varieties 14 by water. 2. Digital Polymerase chain reaction(Droplet-a digital PCR) Primer Small number and gene amplification Garment number [...] possibility mutation (CNVR_13_11, CNVR_8_24, table 1) two garment number response for primer Primer 3 program (http://bioinfo. Ut. Ee/primer3-a 0. 4. 0 /) Through small number-gate. Two garment number possibility mutation marker normalize the number it is generally regulated as an [...] (control) the reference location is desired, a BTF3 gene was used. L of PCR reaction is divided into 2 of 20 micro droplet [...] Droplet-a digital PCR amplification in DNA copy per micro l can be calculates. Digital PCR or RT-a PCR utilized existing garment number variant verification as well as accuracy and higher sensitivity is capable of holding a number of copy (copy resolution) SB. L ddPCR before performing micro gDNA 2 (20ng/micro l), probe (20 pmol/micro l), HindIII hanhyo small total number 20 micro l filling liquid reaction mixture of a silicon wafer is. Said mixture is well each dedicated cartridge QX100 droplet generator which is included in the micro l Generate droplet generation oil 70 (Bio-a Rad, Hercules, CA, USA) to the mounting Droplet produce the substrate. After droplets made less affected by a 96 well PCR plate, DNA Engine Tetrad® 2 Thermal Cycler (Bio-a Rad, Hercules, CA, USA) using amplification reaction chamber. 3. PCR product Analysis and Garment number beNumber variation Calculated QX100 droplet reader (Bio-a Rad, Hercules, CA, USA) is completed PCR 96 well PCR plate is mounted on the, QuantaSoft (Bio-a Rad, Hercules, CA, USA) using droplet number has been confirmed that the number of final DNA variant be checks the signal is desired. Table 1 is for performing primers and probes of the present invention ddPCR is shown disclosed. Result 29 On by selecting a result of bovine chromosome, cattle for later search and retrieval software utilizing 901 CNVSeq possibility mutation of China area along raincoat garment number total number is desired possibility mutation region found. This is 0. Corresponding to 22% value, [...] (0. 63%), It is a hall star (0. 29%) Such as (Choi et al. , 2013, 2014) Variation of garment number compared to be very small numerical been. This cattle area along right indicate that genetically more closely than European health or hypermetropia. (2 Also) dispersed throughout the CNVR gene has been opened. Table 2 is a shadow area along cattle to find a match to said two bovine CNVR sequence by analyzing a major regions are disclosed. Table 3 on bovine chromosome (BTA) identified in revealing the secret key of each of 29 CNVR are disclosed. The various CNVR 1621bp to 161, 921bp into its size and, the size of the average value and each 6147bp 5085bp and been. Subtract cattle and area along the comparison result, of about 53 during the entire CNVR 901. 2% (405 CNVR 2,934,997 bp in) number high number variation is desired, 46. 8% (In 405 CNVR 2,578,343 bp) 24.4 low garment number variant number, when said corresponding to two values, the most CNVR 10kbp hereinafter than pulse, 10 kb hereinafter also been most Ensembl gene (genic provided CNVR) (of Figure 3 A and B). Of CNVGO (Gene ontology) analysis Table 3 reference area along the genome of ontology in cattle and shadow between identified genic provided CNVR exhibit rich genome are disclosed. "Biological attached (Biological adhesion)" and "mobile (locomotion)" appears in the cattle [...] while higher garment number, "metabolism processes (metabolic process)" shadow area along the higher garment number [...] appeared. The aforementioned columns, cattle in China area along side when the number is number of garment number possibility mutation marker difference I 2 be ballistic attack (CNVR_13_11, CNVR_8_24) utilizing a digital PCR on cattle (N=7) (N=7) population number verification experiment number shadow area along China conducting predicting variation is desired. As a result, possibility mutation markers present we shall be in shadow population number [...] CNVR_13_11 garment number area along prevention, prevention of further present in cattle population number [...] 1 has been confirmed. In addition, the biomarker is an area along the shadow population average 0 CNVR_8_24 garment number possibility mutation. 002 Prevention support number has two be transitions, in cattle population average 58. 12 [...] which shows two garment number has been confirmed. The present invention techniques that are directionally-specific content portion more precisely bar, to the party industry in person with skill in the art, such particularly preferred embodiment only the procedure and only aspect, the range of the present invention apparently is the number that are not one will. The substantial range of the present invention defined by appended claim and their equivalent that will. The present invention relates to a composition for identifying varieties of cows including a primer set represented by sequence number 1 and 2, and sequence number 4 and 5, and a method for identifying varieties of cows using the same. COPYRIGHT KIPO 2018 The sequence numbers 3 and 6 can be represented by detecting or amplifying probe including a shadow area along China and cattle number number identifying composition. Back number Back number The sequence numbers 1 and 2 primer set represented; 4 and 5 which enables to cattle and China represented and sequencing number including area along shadow identifying composition. Back number Back number According to Claim 4 composition including cattle and China area along shadow identification kit. Subject nucleic acid sample separation step; said isolated nucleic and IIS, number of marker gene sequence for amplifying primer set 1 and 2 be prevention and sequencing of number 4 and 5 uses a set number of amplified target sequence amplification primer to yield an amplification step; and said amplified gene product by positive number (Copy) confirming including cattle and China area along shadow identification method number number is desired. Back number According to Claim 8, said gene number number identifying step ddPCR (Droplet-a digital PCR) prevention using shadow identification method characterized by analyzing area along cattle and China. According to Claim 8, said gene number number confirming the number 4 anti according to composition is desired, garment number be less than 1 if the rain and dew shadow identification method determining fluctuations area along area along cattle and China.
