CN107586859B - For identifying DNA bar code, primer pair, kit and the method for Shrew Murinus species - Google Patents
For identifying DNA bar code, primer pair, kit and the method for Shrew Murinus species Download PDFInfo
- Publication number
- CN107586859B CN107586859B CN201711058851.1A CN201711058851A CN107586859B CN 107586859 B CN107586859 B CN 107586859B CN 201711058851 A CN201711058851 A CN 201711058851A CN 107586859 B CN107586859 B CN 107586859B
- Authority
- CN
- China
- Prior art keywords
- shrew murinus
- shrew
- murinus
- species
- dna
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention relates to molecular identification technical fields, in particular to a kind of DNA bar code for identifying Shrew Murinus species, primer pair, kit and method.This method comprises: expanding the Cyt b gene segment of Shrew Murinus sample to be measured and being sequenced；Sequence alignment is carried out referring to 14 kinds of DNA bar codes provided by the present invention, if any bar homology in the Cyt b gene segment of the Shrew Murinus sample to be measured and the DNA bar code can determine whether the Shrew Murinus sample to be measured and DNA bar code Shrew Murinus source of species of the same race 96% or more.The present invention is advantageously implemented the quick and precisely identification that China Shrew Murinus belongs to 14 species, shortens qualification time.
Description
Technical field
The present invention relates to molecular identification technical fields, in particular to a kind of for identifying the DNA item of Shrew Murinus species
Shape code, primer pair, kit and method.
Background technique
Shrew Murinus belongs to (Sorex) and is under the jurisdiction of Shrew shape mesh (Soricomorpha) Shrew Murinus section (Soricidae) Shrew Murinus subfamily
(Soricinae) Shrew Murinus, which belongs to, has 16-18 kinds what China was distributed.Shrew Murinus is that a kind of rhynchodaenm point is long, eye is tiny, the original of likeness in form mouse
Beginning mammal, it is closer with the affiliation of mole and hedgehog.Shrew Murinus belongs to the carrying that some species are epidemic hemorrhagic fever viruses
Person, the epidemic hemorrhagic fever virus that different species carry are not quite similar.In external environment and people's contact is close.To its species
Identification is the key that prevention and control Hemorrhagic fever.However, the shape between Shrew Murinus species is more similar, and China point
The Shrew Murinus species of cloth are used for the holotype of morphological classification, none is China research worker acquisition, and this
A little holotypes are all stored in external specimen museum and museum, cause comparative study inconvenient, therefore pass through form
It learns classificating requirement sorter and identification experience is higher.Therefore, it needs to seek a kind of new method, to make up conventional sorting methods
Defect.
DNA bar code technology (DNABarcoding) is analyzed by the DNA sequence dna to a standard target gene,
To quickly and accurately carry out the technology of species identification.In recent years, DNA bar code technology had proved to be having for a row
The bioassay means of effect can not only make strong supplement to conventional identification method, but also because it is more objective, accurate, dash forward
It is broken that experience was depended on unduly in the past, can help to identify species, find novel species and hidden kind, the evolutions of reconstruction species and higher level taxa
Relationship etc..Therefore, if can be very good to belong to China Shrew Murinus by DNA bar code Technology application among the identification that Shrew Murinus belongs to
14 species are quickly and effectively identified.
In view of this, the present invention is specifically proposed.
Summary of the invention
The present invention provides a kind of method for belonging to 14 species DNA bar code Molecular Identifications about China Shrew Murinus, is conducive to reality
Existing China Shrew Murinus belongs to the quick and precisely identification of 14 species, shortens qualification time.
In order to realize above-mentioned purpose of the invention, the following technical scheme is adopted:
One aspect of the present invention is related to a kind of for identifying the DNA bar code of Shrew Murinus species, is selected from SEQ ID NO:1-14
One of shown sequence is a variety of.
DNA bar code of the present invention has the advantages that specific good, amplification efficiency and sequencing success rate are high, is conducive to
The identification of Shrew Murinus species.
According to an aspect of the present invention, the invention further relates to the primer pair for expanding DNA bar code as described above,
Upstream primer and downstream primer are respectively as shown in SEQ ID NO:15 and SEQ ID NO:16.
The primer pair not will form primer dimer and non-specific amplification during PCR amplification, avoid non-purpose product
Influence of the appearance to qualification result, while amplification efficiency is high, and detection signal is strong.
According to an aspect of the present invention, the invention further relates to a kind of Shrew Murinus species identification kit, it includes institutes as above
Primer pair is stated, it is also preferable to include PCR reaction buffer, dNTPs, archaeal dna polymerase, water, sample-loading buffer and DNA molecular amount internal standards
One of or it is a variety of.
According to an aspect of the present invention, the invention further relates to a kind of Shrew Murinus species identification methods, comprising:
The Cyt b gene segment of Shrew Murinus sample to be measured is expanded using kit as described above and is sequenced；
Sequence alignment is carried out referring to DNA bar code as described above, if the Cyt b gene piece of the Shrew Murinus sample to be measured
Section 96% or more, that is, can determine whether the Shrew Murinus sample to be measured and the DNA item with any bar homology in the DNA bar code
Shape code Shrew Murinus source of species of the same race.
This method the professional degree and laboratory condition of technical staff are required it is low, most of R&D institution can be complete
At being conducive to the popularization and promotion of the Shrew Murinus species identification method.
Specific embodiment
One aspect of the present invention is related to a kind of for identifying the DNA bar code of Shrew Murinus species, is selected from SEQ ID NO:1-14
One of shown sequence is a variety of.
The DNA bar code of sequence shown in SEQ ID NO:1-14 is orderly used to the identification of following Shrew Murinus kind:
Big Shrew Murinus Sorex mirabilis, middle Shrew Murinus Sorex caecutiens, Far East Shrew Murinus Sorex isodon, tongue fur
Former Shrew Murinus Sorex tundrensis, long pawl Shrew Murinus Sorex unguiculatus, chestnut tooth Shrew Murinus Sorex daphaenodon,
Thin Shrew Murinus Sorex gracillimus, platycrania Shrew Murinus Sorex roboratus, Ji Shrew Murinus Sorex minutissimus, Yunnan
Shrew Murinus Sorex excelsus, small Shrew Murinus Sorex minutus, back line Shrew Murinus Sorex cylindricaud, small back line Shrew Murinus
Sorex bedfordiae and Tianshan Mountains Shrew Murinus Sorex asper.
According to an aspect of the present invention, the invention further relates to the primer pair for expanding DNA bar code as described above,
Upstream primer and downstream primer are respectively as shown in SEQ ID NO:15 and SEQ ID NO:16.
The primer pair not will form primer dimer and non-specific amplification during PCR amplification, avoid non-purpose product
Influence of the appearance to qualification result, while amplification efficiency is high, and detection signal is strong.
According to an aspect of the present invention, the invention further relates to a kind of Shrew Murinus species identification kit, it includes institutes as above
Primer pair is stated, it is also preferable to include PCR reaction buffer, dNTPs, archaeal dna polymerase, water, sample-loading buffer and DNA molecular amount internal standards
One of or it is a variety of.
Preferably, kit as described above, the archaeal dna polymerase be selected from Taq, Bst, Vent, Phi29, Pfu, Tru,
Tth, Tl1, Tac, Tne, Tma, Tih, Tf1, Pwo, Kod, Sac, Sso, Poc, Pab, Mth, Pho, ES4DNA polymerase,
Klenow segment；
It is highly preferred that the archaeal dna polymerase is Taq archaeal dna polymerase；
It is particularly preferred that the Taq archaeal dna polymerase is high-fidelity Taq archaeal dna polymerase.
Preferably, kit as described above, the wet concentration is from distilled water or deionized water.
According to an aspect of the present invention, the invention further relates to a kind of Shrew Murinus species identification methods, comprising:
The Cyt b gene segment of Shrew Murinus sample to be measured is expanded using kit as described above and is sequenced；
Sequence alignment is carried out referring to DNA bar code as described above, if the Cyt b gene piece of the Shrew Murinus sample to be measured
Section 96% or more, that is, can determine whether the Shrew Murinus sample to be measured and the DNA item with any bar homology in the DNA bar code
Shape code Shrew Murinus source of species of the same race.
Preferably, Shrew Murinus species identification method as described above is thin Shrew Murinus when corresponding kind of the DNA bar code
When Sorex gracillimus, it is desirable that the Cyt b gene segment of sample to be tested and any bar in the DNA bar code are homologous
Property is 96% or more；
It is long pawl Shrew Murinus Sorex unguiculatus or tundra Shrew Murinus Sorex when corresponding kind of the DNA bar code
When tundrensis, it is desirable that the Cyt b gene segment of sample to be tested and any bar homology in the DNA bar code are 96%
More than；
It is big Shrew Murinus Sorex mirabilis, middle Shrew Murinus Sorex when corresponding kind of the DNA bar code
Caecutiens, Far East Shrew Murinus Sorex isodon, chestnut tooth Shrew Murinus Sorex daphaenodon, platycrania Shrew Murinus Sorex
Roboratus, Ji Shrew Murinus Sorex minutissimus, Yunnan Shrew Murinus Sorex excelsus, small Shrew Murinus Sorex
Minutus, back line Shrew Murinus Sorex cylindricaud, small back line Shrew Murinus Sorex bedfordiae or Tianshan Mountains Shrew Murinus Sorex
When asper, it is desirable that the Cyt b gene segment of sample to be tested and any bar homology in the DNA bar code are 98% or more.
Since part Shrew Murinus species distribution is in foreign countries, be different subspecies relationship with domestic distribution, affiliation farther out,
Therefore homology threshold value difference.
The determination of homology threshold value can be by being compared and extracting conservative to 14 kinds of DNA bar codes provided by the present invention
Site, comparing analysis method can be used the progress of this field conventional software, such as Gblocks 0.91b etc., to obtain each DNA item
The homology analysis result of shape code；Threshold value is divided further according to homology analysis result.
Applicant carries out classification and Detection (wherein thin Shrew Murinus 16 to 211 Shrew Murinus using DNA bar code provided herein
A, platycrania Shrew Murinus 2, tundra Shrew Murinus 28, big Shrew Murinus 2, Ji Shrew Murinus 12, chestnut tooth Shrew Murinus 6, Far East Shrew Murinus 44, length
Pawl Shrew Murinus 9, middle Shrew Murinus 54, Yunnan Shrew Murinus 4, small Shrew Murinus 6, back line Shrew Murinus 12, small back line Shrew Murinus 12, Tianshan Mountains Shrew
Murinus 4), and reviewed by multiple means such as source of species, morphology, life habits, it was demonstrated that use the mirror of DNA bar code
Other accuracy rate is up to 100%.
Preferably, Shrew Murinus species identification method as described above expands Shrew Murinus to be measured using kit as described above
When the Cyt b gene segment of sample, annealing temperature is 57 DEG C -59 DEG C；
It is furthermore preferred that annealing temperature is 58 DEG C.
Preferably, Shrew Murinus species identification method as described above expands Shrew Murinus to be measured using kit as described above
When the Cyt b gene segment of sample, PCR response procedures are as follows:
It is furthermore preferred that PCR response procedures are as follows:
Preferably, Shrew Murinus species identification method as described above, in the Cyt b gene segment for expanding Shrew Murinus sample to be measured
Afterwards, before being sequenced further include:
Amplified production is detected with 1%-2% agarose electrophoresis.
Preferably, Shrew Murinus species identification method as described above, the blood of the Shrew Murinus sample to be measured from Shrew Murinus to be measured
Liquid, saliva, sperm, bone or hair.
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the invention.It is not specified in embodiment specific
Condition person carries out according to conventional conditions or manufacturer's recommended conditions.Reagents or instruments used without specified manufacturer is
It can be with conventional products that are commercially available.
Embodiment
Technical scheme is as follows:
Total DNA is extracted from the Shrew Murinus species tissue of acquisition first, carries out PCR amplification Cyt b base using pair of primers
The segment of cause, amplified production is sequenced, and is then carried out sequence analyses and comparison, is established the sequence criteria data of various Shrew Murinus species
Library, on the basis of relatively database D NA, by analyze polymerase chain reaction under given conditions product as a result, from
And reach quickly, the purpose of precise Identification Shrew Murinus species.To the Shrew Murinus sample that needs are identified, its total DNA is extracted, what is given
Under the conditions of, PCR amplification is carried out with the special primer of design, pcr amplification product is determined by agarose gel electrophoresis as a result, then
Sequencing, can identify Shrew Murinus species according to sequence alignment.The molecule mirror for identifying Shrew Murinus species that the present invention designs
Determine method, using following steps:
1, total DNA routinely animal DNA extracting method or blood/cell/tissue genome that Shrew Murinus belongs to animal are extracted
DNA extraction kit carries out DNA extraction, and the DNA concentration of sample is diluted to 0.1 μ g/ μ l-2 μ g/ with the deionized water of sterilizing
μl。
2, amplification of DNA fragments carries out polymerase chain reaction, i.e., is expanded with special primer, primer pair sequence are as follows:
L590:SEQ ID NO:15；
H590:SEQ ID NO:16.
3, PCR product is subjected to agarose gel electrophoresis analysis, detects PCR fragment size using DNAmarker.
4, it the judgement of qualification result: if there is the band of obvious clearly 400bp or so size, and without miscellaneous band, then can send
Biotech firm's sequencing.
5, sequencing result is subjected to manual check and correction, sequence assembly, if carried out with the gene order SEQ ID NO.x same
Source property analysis, if can determine whether described with the homology highest of the gene order SEQ ID NO.x 96% or more
Test serum is that Shrew Murinus belongs to certain source of species.
Specifically, when being thin Shrew Murinus for corresponding kind of the DNA bar code, it is desirable that the Cyt b gene segment of sample to be tested
With any bar homology in the DNA bar code 96% or more；
When being long pawl Shrew Murinus or tundra Shrew Murinus for corresponding kind of the DNA bar code, it is desirable that the Cyt b gene of sample to be tested
Any bar homology in segment and the DNA bar code is 96% or more；
It is big Shrew Murinus, middle Shrew Murinus, Far East Shrew Murinus, chestnut tooth Shrew Murinus, platycrania Shrew Murinus, a Ji when corresponding kind of the DNA bar code
When Shrew Murinus, Yunnan Shrew Murinus, small Shrew Murinus, back line Shrew Murinus, small back line Shrew Murinus or Tianshan Mountains Shrew Murinus, it is desirable that the Cyt b gene of sample to be tested
Any bar homology in segment and the DNA bar code is 98% or more.
Wherein, the DNA bar code method for identifying molecules of 14 kinds of China's Shrew Murinus species is using blood/cell/tissue
Genome DNA extracting reagent kit.
Used archaeal dna polymerase is eTaq high fidelity enzyme.
It carries out using the agarose gel electrophoresis for being 1%-2% when agarose gel electrophoresis analysis.
The band of the 400bp or so size needs to be detected with DNAmaker.
Used DNA sequence dna splicing software includes CodonCode Aligner, Sequencher, Genious, DNA
The softwares such as star.
Compared with traditional Morphological Identification method, the gene order that the present invention obtains, which is advantageously implemented, belongs to China Shrew Murinus
14 species are quickly and effectively identified.
Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention., rather than its limitations；To the greatest extent
Present invention has been described in detail with reference to the aforementioned embodiments for pipe, but those skilled in the art should understand that: its
It is still possible to modify the technical solutions described in the foregoing embodiments, or to some or all of the technical features
It is equivalently replaced；And these are modified or replaceed, various embodiments of the present invention skill that it does not separate the essence of the corresponding technical solution
The range of art scheme.
SEQUENCE LISTING
<110>Mudanjiang Teachers College
<120>for identifying DNA bar code, primer pair, kit and the method for Shrew Murinus species
<160> 16
<170> PatentIn version 3.3
<210> 1
<211> 387
<212> DNA
<213>big Shrew Murinus Sorex mirabilis
<400> 1
atgacaaacc tacgaaaacc cacccactaa taaaaattgt caacagttca tttattgatt 60
tacctgcccc atccaacatt tcatcctgat gaaacttcgg ttccctccta ggcgtctgct 120
taatcatcca aatcctcaca ggactcttcc tagcaataca ctacacatca gacacaataa 180
cagctttttc atcagtcgca cacatttgcc gagacgtaaa ttacggatga ctgattcgct 240
accttcatgc gaatggagca tcaatgttct tcatctgcct gttcctccac gtgggacgag 300
gcctctatta tggatcctac atatatttag aaacatgaaa cattggtgta ctactactat 360
ttacagtaat agccactgcc tttatag 387
<210> 2
<211> 388
<212> DNA
<213>Shrew Murinus Sorex caecutiens in
<400> 2
atgacaaacc tacgaaaaac ccacccatta ataaaaattg ttaacagctc attcattgac 60
ctacctgctc catctaatat ttcatcatga tgaaacttcg gatcccttct aggcgtctgt 120
ctaattatcc aaattcttac aggactcttc ctagctatac actatacatc agacacaata 180
actgccttct cgtcagttac acacatctgc cgagatgtga attacggttg actgatccgc 240
taccttcacg caaacggagc atcaatattc ttcatctgcc tattcctcca cgtaggacga 300
ggcctctatt acgggtccta tatgtattta gaaacatgaa acattggcgt actactgcta 360
ttcgcagtaa tagccactgc ctttatag 388
<210> 3
<211> 387
<212> DNA
<213>Far East Shrew Murinus Sorex isodon
<400> 3
atgacaaacc tacgaaaaac ccacccatta ataaaaatgt taacagttca ttcattgatc 60
tacctgcccc atctaacatc tcatcatgat gaaactttgg ctccctctta ggagtctgct 120
taatcattca aatcctaaca ggactctttc tagcaataca ctacacatca gacacaataa 180
ctgcattttc atcagtaaca cacatctgcc gagacgtaaa ctacggatga ctaatccgct 240
accttcacgc aaatggagca tcaatattct tcatctgcct attcctccac gtcggacgag 300
gcctttatta tgggtcttat atgtacctag aaacatgaaa catcggcgta ctcctcctat 360
tcgcagtaat agctactgcc tttatag 387
<210> 4
<211> 388
<212> DNA
<213>tundra Shrew Murinus Sorex tundrensis
<400> 4
atgacaaacc tacgaaaaac ccacccacta ataaaaattg ttaacagttc attcattgac 60
ctacccgccc cctccaatat ctcatcatga tgaaacttcg gctcccttct aggtgtctgc 120
ttaattattc aaattcttac aggactcttt ctagcaatac actacacatc agacacaatg 180
actgctttct catcagtcac acacatctgc cgagatgtaa actacggatg actaattcga 240
taccttcatg caaacggagc atcaatattc ttcatttgcc tattcctcca cgtcggacga 300
ggtctttact acggctccta catatattta gagacatgaa acatcggtgt attattatta 360
ttcgcagtaa tagccactgc ctttatag 388
<210> 5
<211> 388
<212> DNA
<213>long pawl Shrew Murinus Sorex unguiculatus
<400> 5
atgacaaacc tacgaaaaac ccacccatta ataaaaattg tcaacagttc attcatcgac 60
ttacctgccc catctaacat ctcatcatga tgaaactttg gctccctcct aggtatctgc 120
ttaatcatcc aaatcctaac aggactcttt ttagcaatac actacacatc agacacaata 180
actgcattct catcagttac acacatctgc cgagacgtaa attatggatg actaatccgc 240
taccttcacg caaacggagc atcaatattc ttcatctgcc tattcctcca tgtcggacga 300
ggcctctatt acggatcata catatacctg gaaacatgga acatcggcgt actactgcta 360
tttgcagtaa tagccactgc ctttatag 388
<210> 6
<211> 388
<212> DNA
<213>chestnut tooth Shrew Murinus Sorex daphaenodon
<400> 6
atgacaaacc tacgaaaaac ccacccatta ataaaaattg ttaacagctc attcatcgac 60
ctacccgccc catccaatat ctcatcatga tggaatttcg gctccctcct aggtgtctgc 120
ttaattattc aaatccttac aggactcttt ttagcaatac actacacatc agacacaata 180
actgctttct catcagttac acacatctgt cgagatgtaa attacggatg actaatccga 240
tatcttcatg caaacggagc atcaatattc ttcatttgcc tattcctcca cgtcggacga 300
ggactttact acggatccta tatatactta gagacatgaa acatcggcgt actattacta 360
ttcgcagtaa tagccactgc ctttatag 388
<210> 7
<211> 388
<212> DNA
<213>thin Shrew Murinus Sorex gracillimus
<400> 7
atgacaaacc tacgaaaaac ccacccacta ataaaaattg ttaacagctc attcattgac 60
ctacctgccc catccaatat ctcatcatga tgaaacttcg gctccctcct aggtgtctgc 120
ttaattatcc aaatccttac aggactcttt ctagcaatac actacacatc agacacaata 180
accgctttct catcagtcac acacatctgc cgagatgtga actacggatg actaatccga 240
tatctccacg cgaacggagc atcaatattc tttatctgcc tattcctcca cgtcggacga 300
ggcctctact acggctcata catataccta gaaacatgga atattggcgt actactacta 360
ttcgcagtaa tagccactgc ctttatag 388
<210> 8
<211> 388
<212> DNA
<213>platycrania Shrew Murinus Sorex roboratus
<400> 8
atgacaaacc tacgaaaaac ccacccacta ataaaaattg tgaatagctc attcatcgac 60
ctacccgccc catccaatat ttcatcttga tgaaacttcg gctcccttct aggcatctgc 120
ttaattattc aaatccttac aggactcttc ttagcaatac attatacatc agacacaata 180
actgctttct catcagtcac acacatctgc cgagacgtaa actacggctg actaatccgt 240
tacctgcatg caaacggagc atcaatattc ttcatctgtc tattcctcca cgtcggacga 300
ggcctctact acggatccta catgtactta gaaacatgaa acatcggcgt actactatta 360
ttcgcagtaa tagccaccgc ctttatag 388
<210> 9
<211> 388
<212> DNA
<213>Ji Shrew Murinus Sorex minutissimus
<400> 9
atgacaaacc tacgaaaaac ccacccatta ataaaaattg ttaacagttc atttattgat 60
cttcccgccc catctaacat ctcatcgtga tgaaactttg gctctcttct aggcgtctgc 120
ttaattatcc aaattctcac aggactcttc ttagcaatac attacacatc agacacaata 180
accgccttct catcagttac acatatttgc cgagacgtaa actacggttg actaatccgc 240
tatctccacg caaacggagc atcaatattt ttcatttgcc tgttcctcca cgtcggacga 300
ggcctctact atggctcata catgttttta gagacatgaa acatcggcgt actactatta 360
tttgcagtaa tggccactgc ctttatag 388
<210> 10
<211> 388
<212> DNA
<213>Yunnan Shrew Murinus Sorex excelsus
<400> 10
atgacaaacc tacgaaaaac tcacccatta ataaaaatta ttaacaattc attcattgac 60
ttaccagctc catccaatat ttcatcatga tgaaactttg gctccctcct aggtgtctgc 120
ttaattattc aaatcttaac aggccttttc ctagcaatac actacacatc agacacaata 180
acagcctttt catcagttac tcacatttgt cgagatgtaa actacggttg actaatccgt 240
tacctccacg caaacggagc atcaatattt tttatttgcc tatttctcca cgtcggacga 300
ggcctttact acggatctta catatactta gaaacatgaa atattggtgt actactctta 360
ttcgcagtaa tagccactgc ctttatag 388
<210> 11
<211> 388
<212> DNA
<213>small Shrew Murinus Sorex minutus
<400> 11
atgacaaacc tacgaaaaac ccacccatta ataaaaatcg ttaacagctc atttattgat 60
ttacctgccc catccaatat ttcatcatga tgaaactttg gctccctcct aggtgtctgc 120
ctaattatcc aaatcctcac aggactcttc ctagctatac actacacatc agacacaata 180
actgccttct catcagtcac gcacatctgc cgagacgtaa actacggatg actaatccgc 240
tatctccacg caaacggagc atcaatattc ttcatctgtc tattccttca cgtcggacga 300
ggtctttact acggatccta tatatactta gaaacatgaa acatcggcgt actactacta 360
ttcgcagtaa tagccactgc ctttatag 388
<210> 12
<211> 388
<212> DNA
<213>line Shrew Murinus Sorex cylindricaud is carried on the back
<400> 12
atgacaaacc tacgaaaaac tcacccatta ataaaaatta ttaacagttc atttattgat 60
ttaccagccc catctaacat ttcatcatga tgaaacttcg gctcccttct aggtgtctgc 120
ttaatcatcc aaatcttaac aggccttttt ctagcaatac actatacatc agatacaata 180
acagcctttt catcagttac acacatttgc cgggacgtaa actatggttg actaatccgt 240
taccttcacg caaacggagc atcaatattt tttatttgtc tatttctcca cgttggacga 300
ggcctttact atggatctta catatattta gaaacatgaa acattggtgt tctactctta 360
ttcgcagtaa tagccactgc ctttatag 388
<210> 13
<211> 387
<212> DNA
<213>small back line Shrew Murinus Sorex bedfordiae
<400> 13
atgacaaacc tacgaaaaac ccacccatta ataaaaattg ttaacagttc attcattgac 60
ctaccagctc catccaatat ttcatcatga tgaaactttg gctccctcct aggtgtctgc 120
ttaattatcc aaatcttaac aggccttttt ctagcaatac actacacatc agacacaata 180
acagcctttt catcagtcac acacatttgt cgagacgtaa actatggttg actaatccgt 240
tatctccacg caaacggagc atcaatattt ttcatttgtc tatttctcca cgtcggacga 300
ggcctttact acggacttac atatatttag aaacatgaaa tattggcgta ctactcttat 360
ttgcagtaat agccactgcc tttatag 387
<210> 14
<211> 388
<212> DNA
<213>Tianshan Mountains Shrew Murinus Sorex asper
<400> 14
atgacaaacc tgcgaaaaac ccatccacta ataaaaattg ttaatagttc attcatcgac 60
ctacctgccc cctctaacat ctcatcatga tgaaatttcg gctcccttct aggtgtctgc 120
ttaattattc aaattcttac aggactcttt ttagcaatac actacacatc agacacaata 180
actgctttct catcagtcac acacatctgc cgagatgtaa actacggatg gctaattcga 240
tatcttcatg caaacggagc atcaatattc tttatttgcc tattcctcca cgtcggacga 300
ggtctttact acggctccta catatatcta gaaacatgaa acatcggtgt attattatta 360
ttcgcagtaa tagccactgc ctttatag 388
<210> 15
<211> 23
<212> DNA
<213>artificial sequence
<400> 15
agcactgaaa atgcttagat gag 23
<210> 16
<211> 23
<212> DNA
<213>artificial sequence
<223>r=g or a.
<400> 16
ctataaaggc rgtggctatt act 23
Claims (1)
1. for identifying the DNA bar code combination product of Shrew Murinus species comprising sequence shown in SEQ ID NO:1-14.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711058851.1A CN107586859B (en) | 2017-11-01 | 2017-11-01 | For identifying DNA bar code, primer pair, kit and the method for Shrew Murinus species |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711058851.1A CN107586859B (en) | 2017-11-01 | 2017-11-01 | For identifying DNA bar code, primer pair, kit and the method for Shrew Murinus species |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107586859A CN107586859A (en) | 2018-01-16 |
| CN107586859B true CN107586859B (en) | 2019-01-29 |
Family
ID=61043427
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711058851.1A Active CN107586859B (en) | 2017-11-01 | 2017-11-01 | For identifying DNA bar code, primer pair, kit and the method for Shrew Murinus species |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107586859B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105349658A (en) * | 2015-11-26 | 2016-02-24 | 四川农业大学 | Molecular biological method for quickly identifying Budorcas taxicolor |
| CN105483227A (en) * | 2015-12-22 | 2016-04-13 | 华大(镇江)水产科技产业有限公司 | Sinilabeo rendahli DNA (deoxyribonucleic acid) bar code standard detection gene and application thereof |
-
2017
- 2017-11-01 CN CN201711058851.1A patent/CN107586859B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105349658A (en) * | 2015-11-26 | 2016-02-24 | 四川农业大学 | Molecular biological method for quickly identifying Budorcas taxicolor |
| CN105483227A (en) * | 2015-12-22 | 2016-04-13 | 华大(镇江)水产科技产业有限公司 | Sinilabeo rendahli DNA (deoxyribonucleic acid) bar code standard detection gene and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| DNA条形码技术在小型兽类鉴定中的探索：以甘肃莲花山为例;何锴 等;《生物多样性》;20131231;第21卷(第2期);第199页左栏第2.3-2.4部分，右栏倒数第2段至第200页左栏第1段,第203页第4.3部分 |
| Evolution of the cytochrome b gene of mammals;David M.Irwin;《J Mol evol》;19911231;第32卷;128-144 |
| 巴郎沟小型兽类区系及群落多样性研究与Cytb辅助鉴定未知鼩鼱;侯全芬;《中国优秀硕士学位论文全文数据库》;20130815(第8期);第二章第25-33页 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107586859A (en) | 2018-01-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111394520B (en) | Primer group for detecting new coronavirus based on RT-LAMP technology and detection kit | |
| CN108060257B (en) | Primer composition for detecting pythium androsaceus based on loop-mediated isothermal amplification technology and detection method thereof | |
| US11795451B2 (en) | Primer for next generation sequencer and a method for producing the same, a DNA library obtained through the use of a primer for next generation sequencer and a method for producing the same, and a DNA analyzing method using a DNA library | |
| CN107142334B (en) | RT-LAMP detection method of mulberry vein banding virus, primer group, kit and application thereof | |
| CN103966228B (en) | A kind of Rh blood group DEL type RHD93T > A allelotrope and detection method thereof | |
| ES2719412T3 (en) | Control of NGS systems and methods that include them | |
| WO2011146942A1 (en) | Methods and kits to analyze microrna by nucleic acid sequencing | |
| CN109371146A (en) | SNP marker, primer pair, detection kit and its application of the more thoracic vertebrae number characters of sheep | |
| CN103937806B (en) | A kind of Rh blood group DEL type RHD838G > A allelotrope and detection method thereof | |
| CN107586859B (en) | For identifying DNA bar code, primer pair, kit and the method for Shrew Murinus species | |
| KR20190026403A (en) | Primers for Analyzing Porcine Epidemic Diarrhea Virus Genotype and Uses Thereof | |
| CN108998576A (en) | For detect the specific primer of spring viremia of carp virus to, probe, detection kit | |
| CN109055565B (en) | DNA bar code of Moina mongolica and application thereof | |
| CN112430651A (en) | Detection method for integrity of FFPE sample | |
| CN112795662A (en) | Identification method of barbel grahami and application thereof | |
| JP6853523B2 (en) | PCR using helicase | |
| CN112048573A (en) | RPA primer and kit for detecting cotton leaf curl virus, detection method and application thereof | |
| CN114958875B (en) | Screening and application of reference gene metG of citrus yellow-long pathogen | |
| CN109182599A (en) | For detect the specific primer of grass carp hemorrhage virus to, probe, detection kit | |
| KR101735762B1 (en) | Prediction method for swine fecundity using gene expression profile | |
| KR102279787B1 (en) | Specific primer set for detecting ChiVMV and uses thereof | |
| CN111808975B (en) | Deer species source identification primer pair of deer antler product and identification method thereof | |
| CN114107539B (en) | LAMP primer composition for detecting pythium delicicola and application thereof | |
| KR102147327B1 (en) | A composition for detecting Ganoderma microorganism and diagnosing basal stem rot and a method using the same | |
| KR100376770B1 (en) | Rapid Multiplex Genotyping Kit System for the detection of Y-chromosome STRs |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |