KR20140016538A - Molecular marker and analysis method for identifiction of river drainage system of gobiobotia macrocephala - Google Patents
Molecular marker and analysis method for identifiction of river drainage system of gobiobotia macrocephala Download PDFInfo
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Abstract
Description
본 발명은 금강 꾸구리와 한강 꾸구리의 집단 식별을 위한 마커와 상기 마커를 이용한 금강 꾸구리와 한강 꾸구리의 집단 식별 방법에 관한 것이다.The present invention relates to a marker for collective identification of Geumganganggwang and Hanganggukri, and a method for collecting identification of Geumganggukri and Hanganggurri using the marker.
본 연구종인 꾸구리(Gobiobotia macrocephala)는 우리나라 고유종으로 모래무지아과의 꾸구리속(Gobiobotia)에 속하며 소형 저서성 어류로 금강집단, 한강집단으로 나뉘고 물살이 빠르고 자갈과 돌이 많이 깔린 중상류 지역에 서식한다. 최근 환경오염과 댐건설, 하천개발 등으로 인해 개체수가 급감하고 있어 환경부는 2005년 멸종위기 야생동·식물 Ⅱ급으로 지정하여 법적 보호를 하고 있다.This species, Gobiobotia macrocephala , is endemic to Korea and belongs to the genus Gobiobotia. It is a small benthic fish divided into Geumgang group and Han river group. In recent years, due to environmental pollution, dam construction, and river development, the number of the population has been declining, and the Ministry of Environment has designated it as the endangered wildlife,
우리나라의 지형적인 특성 및 독립성이 높은 담수계의 특징으로 인하여 동일 종 내에서도 서식하는 수계에 따라 집단 간 유전적으로 높은 분화가 나타나고 있음이 보고되고 있다. 최근 생물이 가지고 있는 유전자의 염기서열을 분석하여 생물의 계통을 추론하는 분자계통학은 가장 강력한 생물 계통 추적법의 하나로 인정되어 널리 활용되고 있다. 이러한 계통분석법의 성공에 힘입어 최근에는 종의 분류에 분자계통분석법을 적용하려는 DNA taxonomy란 개념이 제기되어 관련 연구자들로부터 많은 주목을 받고 있다.It has been reported that due to the characteristics of the freshwater system, which has high geographical characteristics and independence of Korea, highly differentiated genetically between groups according to the water system inhabiting the same species. Recently, molecular genetics, which infer the biological system by analyzing the nucleotide sequence of a gene possessed by a living organism, is recognized as one of the most powerful biological system tracking methods and widely used. Due to the success of this systematic analysis method, the concept of DNA taxonomy to apply the molecular phylogenetic analysis to the classification of species has recently been attracting much attention from related researchers.
선진국의 경우 고유의 어종에 관한 정부의 적극적인 관심으로 일본의 경우 우리나라 수입산 및 자국산 고등어의 rapid PCR-RFLP method의 개발로 연안산과 대서양산의 식별, 수입산 갈치의 mtDNA 분석 등을 통해 자국 내 수입어류에 관한 원산지 식별을 이룬바 있으며, 미국은 농무성 산하 FAS (Foreign agricultural service)에서 정보관리를 하는 등의 정부 기관 운영 하에 원산지 식별 기술에 노력하고 있다. 물론 국내의 경우도 연어 및 홍어류의 종 동정을 위한 DNA chip 기술개발, 연어과와 숭어과 어류의 mtDNA에 의한 분자계통학적 연구, RAPD, AFLP 및 microsatellite를 이용해 다양한 종의 유전다양성 분석 등의 연구는 진행되고 있다.In the case of developed countries, the government's active interest in the native fish species has led to the development of the rapid PCR-RFLP method of imported and exported mackerel in Korea. In Japan, identification of the marine and Atlantic mountains and mtDNA analysis of the imported hawk And the United States is working on the identification technology of origin under the operation of government agencies such as information management in FAS (Foreign agricultural service) under the Ministry of Agriculture. Of course, in Korea, development of DNA chip technology for identification of salmon and red sea bream species, study of molecular diversity by mtDNA of salmon, mullet and fish, genetic diversity analysis of various species using RAPD, AFLP and microsatellite .
또한 COl 영역 분석 결과에서 생성된 특이 DNA 밴드의 sequencing 과정을 거쳐 SCAR 마커 프라이머를 제작하여 PCR을 수행하는 것이 필요하다. 이 방법은 개체 간 차이를 보이는 DNA 염기서열로부터 DNA를 안정되게 증폭시킬 수 있는 한 쌍의 프라이머를 제작하여 PCR 과정을 수행하기 때문에 특이도가 향상된 결과를 얻을 수 있는 장점이 있다. In addition, it is necessary to perform PCR by constructing SCAR marker primers through sequencing of specific DNA bands generated from COl region analysis results. This method has the advantage of achieving improved specificity because PCR is performed by preparing a pair of primers capable of stably amplifying DNA from DNA sequences showing differences between individuals.
본 발명은 상기한 바와 같이 금강 꾸구리와 한강 꾸구리의 방류시 혼입으로 인한 문제를 해결하기 위해 제안된 것으로, 본 발명은 금강 및 한강 꾸구리의 상호간에 집단 식별이 가능한 마커 및 상기 마커를 이용한 금강 꾸구리와 한강 꾸구리의 집단 식별 방법을 제공하는 것을 목적으로 한다. As described above, the present invention has been proposed in order to solve the problems caused by mixing in the discharge of the Geumganggunggi and the Hangangguri, and the present invention is applicable to the case where the marker can be collectively identified among the Geumgang and Hangangguri, It is aimed to provide a group identification method of Hangang Guquri.
상기와 같은 목적을 달성하기 위한 본 발명의 꾸구리 mitochondrial DNA의 COI영역의 PCR 증폭용 프라이머는 서열번호 1 또는 서열번호 2의 염기서열을 포함하는 것을 특징으로 한다. Primer for PCR amplification of the COI region of the coo mitochondrial DNA of the present invention for achieving the above object is characterized in that it comprises a nucleotide sequence of SEQ ID NO: 1 or SEQ ID NO: 2.
또한, 본 발명의 일 실시예에 따른 꾸구리 수계 분석용 SCAR 마커는 서열번호 3 내지 서열번호 6 중 어느 한 프라이머를 포함하는 것을 특징으로 한다. In addition, the SCAR marker for analysis of a cooing tree according to an embodiment of the present invention is characterized in that it comprises any one of SEQ ID NO: 3 to SEQ ID NO: 6.
또한, 본 발명의 일 실시예에 따른 금강에 서식하는 꾸구리 수계 분석용 SCAR 마커는 서열번호 5 또는 서열번호 6의 프라이머를 포함하는 것을 특징으로 한다. In addition, the SCAR marker for coo water system analysis inhabiting Geumgang according to an embodiment of the present invention is characterized in that it comprises a primer of SEQ ID NO: 5 or SEQ ID NO: 6.
또한, 본 발명의 바람직한 일 실시예에 따른 꾸구리 수계 분석방법은 꾸구리의 genomic DNA를 분리하는 단계; 상기 분리된 genomic DNA를 주형으로 하여 서열번호 1 또는 서열번호 2의 돌상어 mitochondrial DNA의 COI영역의 PCR 증폭용 프라이머로 증폭시키는 단계; 상기 증폭된 산물을 전기영동하는 단계; 및 상기 전기영동 후 증폭된 산물이 2개 이상이면 금강 꾸구리, 1개만 존재하면 한강 꾸구리로 식별하는 단계를 포함하는 것을 특징으로 한다. In addition, according to one preferred embodiment of the present invention, a method for analyzing a cooing tree is characterized by separating genomic DNA of coo; Amplifying the isolated genomic DNA with a primer for PCR amplification of the COI region of the shark mitochondrial DNA of SEQ ID NO: 1 or SEQ ID NO: 2; Electrophoresis of the amplified product; And if the product amplified after the electrophoresis is two or more, the step of identifying the Geumgang copuri, and if only one is present as a Hanguri copuri.
본 발명에 따른 금강과 한강 꾸구리의 수계 식별용 마커는 분자생물학적인 기술을 이용하여 개발된 마커로서, 기존의 형태적 형질로 구분할 수 없는 꾸구리를 집단별로 구분이 가능하다. 또한, 단 한번의 PCR 증폭 및 전기영동을 통하여 금강집단과 한강집단 꾸구리를 식별할 수 있는 기술로서, 신속하고 간편하며 정확성이 높아 집단 식별의 효율성과 실용성을 극대화할 수 있는 장점이 있다.According to the present invention, the markers for identification of water in the Geum River and Hangang Ridge are developed using molecular biology techniques, and can be classified into groups that can not be classified into existing morphological traits. In addition, it is a technique that can distinguish between the Kumgang group and the Han river group through a single PCR amplification and electrophoresis. It is fast, simple and highly accurate, and has the advantage of maximizing group identification efficiency and practicality.
본 발명에 따라 신속하고 정확한 꾸구리의 표시가 이루어지면 방류시 혼입을 막을수 있고, 우리나라의 고유 어종에 대한 정확한 집단 동정 및 보호를 위한 기초적인 개발이 될 것이다. According to the present invention, if quick and precise display is performed, it will prevent mixing at the time of discharge, and it will be a basic development for accurate identification and protection of the native species of Korea.
도 1은 본 발명의 Mitochondrial DNA COl 영역에서 검출한 꾸구리 SCAR 마커의 Multiplex PCR 산물의 전기영동 사진(GR : 금강수계 집단, HR : 한강수계 집단)이다.
도 2는 각 수계 집단별 SCAR 마커를 이용한 검증 결과를 나타낸 것이다.
도 3은 꾸구리 미토콘드리아 영역 중 COI 영역에서 나타난 변이 사이트를 나타낸 것이다. 1 is a mitochondrial DNA COl of the present invention Electrophoresis photographs of multiplex PCR products of coo SCAR markers detected in the area (GR: Geum River Group, HR: Han River Group).
FIG. 2 shows the results of the verification using SCAR markers for each water group.
FIG. 3 shows mutation sites in the COI region of the conger eel mitochondrial region.
본 발명은 꾸구리의 미토콘드리아 DNA의 cytochrome c oxidae I (COI)영역의 집단 특이 변이 사이트를 이용하여 SCAR(sequence characterized amplified region) Marker 프라이머를 제작하고, 이것을 통해 금강 꾸구리 집단과 한강 꾸구리 집단을 구분할 수 있는 마커를 개발하였다. 본 기술은 신속하게 꾸구리의 이입개체를 판별할 수 있는 기술로서 방류시 개체 혼합유입을 막으며, 자연개체군의 인위적 이입에 대한 관리방안 마련을 위해 개발되었다.In the present invention, a SCAR (Sequence characterized amplified region) marker primer is constructed using the population-specific mutation site of the cytochrome c oxidae I ( CO I) region of the mitochondrial DNA of conger eel, Developed a marker. This technology is a technology that can quickly identify the entrances of the entomologists, and it was developed to prevent the influx of entities at the time of discharge and to prepare a management plan for the intrusive introduction of natural populations.
본 실험에 이용된 마커의 경우 신속하고 간편하며 정확성이 높아 방류시의 효율성과 실용성을 극대화 할수 있으며 멸종위기종 꾸구리 방류시나 혼합되었을 시 두 집단을 나눌 수 있는 마커로 쓰일 수 있다.The marker used in this experiment can be used as a marker to divide the two groups when discharged or mixed with endangered species, because it is quick, simple and highly accurate, thus maximizing efficiency and practicality at the time of discharge.
이하, 실시예를 통해 본 발명을 상세히 설명하기로 한다. 하기에 제시된 실시예는 예시적인 것으로 이 분야에서 통상의 지식을 가지는 자는 본 발명의 기술적 사상을 벗어나지 않는 범위에서 제시된 실시예에 대한 다양한 변형 및 수정 발명을 만들 수 있을 것이다. 이러한 변형 및 수정 발명에 의하여 본 발명의 범위는 제한되지 않는다.Hereinafter, the present invention will be described in detail with reference to examples. It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the invention. The scope of the present invention is not limited by these variations and modifications.
다음의 실시 예에 따라 본 발명을 상세히 설명한다.
The present invention will be described in detail with reference to the following examples.
[[ 실시예Example 1 : One : 꾸구리Prickly 지느러미 조직으로부터 From the fin tissue genomicgenomic DNADNA 추출] extraction]
본 발명에 사용된 꾸구리의 genomic DNA는 한강수계인 홍천강, 섬강, 임진강 3개 집단과 금강 1개 집단을 사용하였으며, 추출은 개체의 꼬리지느러미를 약 1 cm2 정도 절단하여 페놀 추출법을 이용하여 추출하였다.
The genomic DNAs used in the present invention were the Hongcheon River, the Sumgang River, the Imjin River, and the Geum River, and the extraction of the genomic DNA was carried out to about 1 cm 2 And extracted with phenol extraction method.
[[ 실시예Example 2 : 2 : PCRPCR 을 통한 through 꾸구리의Prickly 유전자의 증폭 및 염기서열 비교분석] Amplification of genes and comparison of base sequences]
(1) PCR을 통한 꾸구리의 개체별 유전자 증폭 (1) Gene amplification of each individual by PCR
꾸구리의 집단 특이 SCAR maker를 검출하기 위해 각 개체의 genomic DNA를 이용하여 mitochondrial DNA의 COI영역을 PCR 증폭하였다. 프라이머 제작은 GenBank에서 이용 가능한 어류종들의 유전정보를 내려받아 사용하였다. 이들 유전자 정보는 BioEdit 7.0.1의 ClustalW를 이용하여 다중서열정렬을 수행한 후, 보존성이 높은 부분의 염기서열 정보를 바탕으로 제작하였다. 본 발명에 이용된 프라이머 염기서열은 표 1에 제시하였다.The genomic DNA of each individual was used to PCR amplify the COI region of mitochondrial DNA to detect the population - specific SCAR maker. Primer production was used to download genetic information of available species in GenBank. These gene information were constructed based on sequence information of highly conserved regions after performing multiple sequence alignment using ClustalW of BioEdit 7.0.1. The primer base sequences used in the present invention are shown in Table 1.
PCR 반응은 20 ㎕ 용적의 AccuPower PCR Premix Kit (Bioneer, Korea)에 유전체 100 ng DNA 2㎕와 5 pmole의 각 프라이머 1 ㎕를 넣은 후, 94℃에서 4분 간 초기 열변성 반응을 시킨 후, 94℃ 30초, 50℃ 30초, 72℃ 1분의 순환 반응을 40회 실시하였다. 최종적으로 72℃ 7분간의 신장 반응을 시킨 후, PCR 반응의 성공 여부를 GelRed (Invitrogen, USA)로 염색된 1.5% agarose gel에서 전기 영동하여 확인하였다. PCR was carried out by adding 2 μl of a genomic DNA of 100 ng and 1 μl of each primer to 20 μl of AccuPower PCR Premix Kit (Bioneer, Korea), followed by initial thermal denaturation at 94 ° C for 4 minutes, followed by 94 Circulation was repeated 40 times at 30 ° C for 30 seconds, at 50 ° C for 30 seconds, and at 72 ° C for 1 minute. After a final extension at 72 ° C for 7 minutes, the success of the PCR reaction was confirmed by electrophoresis on 1.5% agarose gel stained with GelRed (Invitrogen, USA).
(2) Direct_sequencing을 통한 꾸구리 개체별 염기서열 비교분석(2) Comparison of nucleotide sequence analysis by direct sequencing
PCR증폭된 개체의 유전자 염기서열 비교분석을 수행하기 위하여 PCR 산물을 대상으로 AccuPrep PCR Purification Kit (Bioneer)를 이용하여 정제한 후, 자동염기서열분석기(ABI 3700; Perkin Elmer)를 이용하여 염기서열을 결정 및 분석하였다.
PCR products were purified using the AccuPrep PCR Purification Kit (Bioneer), and sequenced using an automated sequencer (ABI 3700; Perkin Elmer). And analyzed.
[[ 실시예Example 3 :미토콘드리아 영역을 통한 3: Through the mitochondrial region 꾸구리의Prickly 집단 특이 Group specific DNADNA 영역 확인 및 Zone verification and 프라이머primer 제작] making]
PCR 결과 COI영역 1551bp모두가 증폭이 확인 되었고 증폭된 COl영역 중 보존성이 높은 염기서열에서 꾸구리 집단 마커 CFM과 CRM을 개발하였고, 금강 꾸구리 집단의 haplotype으로 판단되는 126bp, 129bp, 177bp, 234bp, 261bp, 264bp, 285bp, 291bp, 318bp, 393bp, 399bp, 408bp, 441bp, 543bp, 588bp, 603bp, 663bp, 741bp, 765bp, 786bp, 795bp, 807bp, 816bp, 861bp, 882bp, 885bp, 888bp, 900bp, 954bp, 990bp, 1020bp, 1041bp, 1077bp, 1125bp, 1158bp, 1195bp, 1244bp, 1374bp 염기가 발견되어 본 실험자는 금강 꾸구리 집단의 haplotype으로 판단되는 COI 영역 중 885bp, 888bp, 900bp영역을 이용하여 금강 꾸구리 집단 특이마커 GRM을 개발하였다(도 3 참조). As a result of PCR, amplification of all 1551bp of COI region was developed, and we developed coupe group markers CFM and CRM from the highly conserved nucleotide sequence of the amplified COl region, and 126bp, 129bp, 177bp, 234bp, 261bp, 264bp, 285bp, 291bp, 318bp, 393bp, 399bp, 408bp, 441bp, 543bp, 588bp, 603bp, 663bp, 741bp, 765bp, 786bp, 795bp, 807bp, 816bp, 861bp, 882bp, 885bp, 888bp, 900bp, 954bp, 990bp 1020bp, 1041bp, 1077bp, 1125bp, 1158bp, 1195bp, 1244bp, and 1374bp bases were found, and the present inventors developed 885bp, 888bp, and 900bp regions of the Geum coupe population-specific marker GRM using the COI region determined to be the haplotype of the Geum copper population. (See FIG. 3).
본 발명에 이용된 집단 식별 SCAR 마커 프라이머 염기서열을 표 2에 제시하였으며 꾸구리 집단의 SCAR 마커에 대한 Multiplex PCR을 실시 하였다.The nucleotide sequences of the collective identification SCAR marker primers used in the present invention are shown in Table 2, and Multiplex PCR was performed on the SCAR markers of the convalescent group.
꾸구리 집단 식별 SCAR 마커의 PCR반응은 20 ㎕ 용적의 AccuPower Hotstart PCR Premix Kit (Bioneer, Korea)에 유전체 100 ng DNA 1㎕와 10 pmole의 CFM 프라이머 2㎕, CRM 프라이머 1.5㎕, GRM 프라이머 0.3㎕ 넣은 후, 94℃에서 4분간 초기 열변성 반응을 시킨 후, 94℃ 30초, 56℃ 30초, 72℃ 1분의 순환 반응을 35회 실시하였다. 최종적으로 72℃에서 7분간의 신장 반응을 시킨 후, PCR 반응의 성공 여부를 GelRed (Invitrogen, USA)로 염색된 2.0% agarose gel에서 전기영동하여 확인하였다.PCR reaction of conidial-group identification SCAR marker was carried out by adding 1 μl of genomic DNA of 100 ng DNA, 20 μl volume of AccuPower Hotstart PCR Premix Kit (Bioneer, Korea), 2 μl of 10 pmole CFM primer, 1.5 μl of CRM primer and 0.3 μl of GRM primer , An initial thermal denaturation reaction was carried out at 94 ° C for 4 minutes, and then a circulation reaction was performed at 94 ° C for 30 seconds, 56 ° C for 30 seconds and 72 ° C for 1 minute for 35 times. After a final extension at 72 ° C for 7 minutes, the success of the PCR reaction was confirmed by electrophoresis on 2.0% agarose gel stained with GelRed (Invitrogen, USA).
그 결과 한강 꾸구리 집단에서는 금강 꾸구리 집단에서도 동일하게 나타나는 밴드가 나타났고 금강 꾸구리 집단에서는 한강 꾸구리 집단에서도 나타나는 밴드뿐만 아니라 금강 꾸구리 집단에서만 나타나는 금강 꾸구리 특이 밴드도 나타났다.
As a result, the same band appeared in the Yangguang congregation group as well as in the Yangguang congregation group.
Gobiobotia macrocephala
(CFM-CRM)Common
Gobiobotia macrocephala
(CFM-CRM)
Gobiobotia macrocephala
(CFM-GRM)Geum River
Gobiobotia macrocephala
(CFM-GRM)
<110> Foundation of Sunchunhyang University and Industry Cooperation <120> Molecular Marker for Identifiction of Water System of Gobiobotia macrocephala and the Method of Identifying the Water System of Gobiobotia macrocephala Using the Same <130> P-2012-00011 <160> 6 <170> KopatentIn 2.0 <210> 1 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> PCR primer COI F3 <400> 1 gatcctrcaa aytcytagtt aacagcta 28 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> PCR primer COI R3 <400> 2 tgraatccta rttghgwggg 20 <210> 3 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> SCAR Marker pimer foward CRM-CRM <400> 3 ttggtaccct ytatcttgta tttg 24 <210> 4 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> SCAR Marker pimer reverse CRM-CRM <400> 4 ctcctcgaat gtgtggtat 19 <210> 5 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> SCAR Marker pimer foward CRM-GRM <400> 5 ttggtaccct ytatcttgta tttg 24 <210> 6 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> SCAR Marker pimer reverse CRM-GRM <400> 6 gtgtctacgt ctatcccg 18 <110> Foundation of Sunchunhyang University and Industry Cooperation <120> Molecular Marker for Identification of Water System of Gobiobotia macrocephala and the Method of Identifying the Water System Gobiobotia macrocephala Using the Same <130> P-2012-00011 <160> 6 <170> Kopatentin 2.0 <210> 1 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> PCR primer COI F3 <400> 1 gatcctrcaa aytcytagtt aacagcta 28 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> PCR primer COI R3 <400> 2 tgraatccta rttghgwggg 20 <210> 3 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> SCAR Marker pimer foward CRM-CRM <400> 3 ttggtaccct ytatcttgta tttg 24 <210> 4 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> SCAR Marker pimer reverse CRM-CRM <400> 4 ctcctcgaat gtgtggtat 19 <210> 5 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> SCAR Marker pimer foward CRM-GRM <400> 5 ttggtaccct ytatcttgta tttg 24 <210> 6 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> SCAR Marker pimer reverse CRM-GRM <400> 6 gtgtctacgt ctatcccg 18
Claims (4)
상기 분리된 genomic DNA를 주형으로 하여 서열번호 1 또는 서열번호 2의 돌상어 mitochondrial DNA의 COI영역의 PCR 증폭용 프라이머로 증폭시키는 단계;
상기 증폭된 산물을 전기영동하는 단계; 및
상기 전기영동 후 증폭된 산물이 2개 이상이면 금강 꾸구리, 1개만 존재하면 한강 꾸구리로 식별하는 단계;
를 포함하는 것을 특징으로 하는 꾸구리 수계 분석방법. Isolating genomic DNA from coo;
Amplifying the isolated genomic DNA with a primer for PCR amplification of the COI region of the shark mitochondrial DNA of SEQ ID NO: 1 or SEQ ID NO: 2;
Electrophoresing the amplified product; And
Identifying two or more amplified products after the electrophoresis as Geum-Gu-copper, and if only one is present, as Han-Gang-Cu;
Cubic water system analysis method comprising a.
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| KR101499693B1 (en) * | 2014-11-04 | 2015-03-16 | 대한민국 | Single Nucleotide Polymorphic DNA markers for discriminating habitat of the Korean striped bitterling, Acheilognathus yamatsutae and a method using the same |
| CN111500745A (en) * | 2020-04-29 | 2020-08-07 | 北京林业大学 | Long fragment amplification primer for eliminating interference of mitochondrion pseudogene of dendroctonus valens |
| CN116926210A (en) * | 2023-08-30 | 2023-10-24 | 江西省水产科学研究所(江西省鄱阳湖渔业研究中心、江西省渔业资源生态环境监测中心) | Specific COI (chip on insulator) primers and application thereof in rapid identification of rana spinosa |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101499693B1 (en) * | 2014-11-04 | 2015-03-16 | 대한민국 | Single Nucleotide Polymorphic DNA markers for discriminating habitat of the Korean striped bitterling, Acheilognathus yamatsutae and a method using the same |
| CN111500745A (en) * | 2020-04-29 | 2020-08-07 | 北京林业大学 | Long fragment amplification primer for eliminating interference of mitochondrion pseudogene of dendroctonus valens |
| CN111500745B (en) * | 2020-04-29 | 2022-04-26 | 北京林业大学 | Long fragment amplification primer for eliminating interference of mitochondrion pseudogene of dendroctonus valens |
| CN116926210A (en) * | 2023-08-30 | 2023-10-24 | 江西省水产科学研究所(江西省鄱阳湖渔业研究中心、江西省渔业资源生态环境监测中心) | Specific COI (chip on insulator) primers and application thereof in rapid identification of rana spinosa |
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