CN1965664A - Method for determining if common cow milk has been doped in yak milk product and primer dedicated therefor - Google Patents
Method for determining if common cow milk has been doped in yak milk product and primer dedicated therefor Download PDFInfo
- Publication number
- CN1965664A CN1965664A CN 200610114329 CN200610114329A CN1965664A CN 1965664 A CN1965664 A CN 1965664A CN 200610114329 CN200610114329 CN 200610114329 CN 200610114329 A CN200610114329 A CN 200610114329A CN 1965664 A CN1965664 A CN 1965664A
- Authority
- CN
- China
- Prior art keywords
- yak
- sequence
- milk
- dairy produce
- 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.)
- Granted
Links
Images
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to a method for checking the common milk in yak milk, wherein said method comprises that: (1), using the DNA extracted from the yak milk as template, using one couple of inductors whose nucleotide sequences are the sequence 1 and sequence 2 of sequence list to process PCR reaction; (2), checking if the expand product of PCR has DNA band, if there is, the yak milk is doped with common milk. The invention has high accuracy.
Description
Technical field
The present invention relates to a kind of method and primer special thereof of differentiating the ordinary milk that whether mixes in the yak dairy produce.
Background technology
Yak be in the domestic animal the extremely limited rare domestic animal of geographic range it, yak milk and dairy produce thereof claim to show with its rare and high nutritive value with respect to ordinary milk.Therefore some illegal retailers just occurred, in yak milk, mixed ordinary milk and therefrom seek higher profit, made consumer's interests be subjected to great infringement.
Yak mainly lives in plateau, high mountain, alpestrine cold half humid region of height above sea level 2000-4500m.The China yak accounts for more than 90% of world yak, mainly is distributed in Qinghai-Tibet central area and east margin area thereof.The distribution of yak is extremely limited, and as rare domestic animal kind, yak can provide products such as breast, meat, fat, hair, suede, skin for people.In these products particularly the breast processing, utilize the form kind various.Because being of high nutritive value of yak dairy products, the output rareness, so its market price is than high many of common dairy product.
Galactopoiesis season of yak after annual calving in May to November, peak of lactation 7, August, the galactopoiesis phase is about 150 days.The milk production of yak is lower, and the milk production in 150 days milking phases is about 350kg, about average milking every day amount 1.0-2.5kg.The highest with Xinjiang Ba Zhou yak in the kind of yak each place, 2.5kg/ days.And common at home milk cow is divided into 305 days lactation period, and the average output of milk is 4000kg, and average milking every day amount is 13.5kg.From nutritional labeling relatively, the nutrient composition content of yak milk is than high many of ordinary milk, and Fig. 1 has compared the difference on the nutrient composition content between common cow's milk, colostrum and yak emulsion, the colostrum.From data as can be seen, two main features of yak milk: be of high nutritive value, and rare.
The length of mammal mitochondrial DNA (mtDNA) is about 16kb, and the length of human mtDNA is 16569bp; MtDNA is the closed hoop structure, is positioned at mitochondrial matrix, and thousands of copy mitochondrias (different and different according to cell type) are arranged in each cell.It is double-stranded that mitochondrial DNA is, a heavy chain (H) that is rich in guanine, a light chain (L) that is rich in cytimidine.12 polypeptide chains of heavy chain coding, 14 tRNA and all rRNA; Polypeptide chain of light chain coding, 8 tRNA.The gene inside of mitochondria coding does not have introne, and all coded sequences closely link to each other in the gene.
Only noncoding region is D-loop district (Displacement loop) in the mitochondrial DNA, should the long 1121bp in zone in human mtdna, and this zone includes heavy chain replication origin (O
H) and light chain, the transcripting starting subarea of heavy chain.The replication initiation of mitochondrial DNA duplicates and occur in heavy chain replication origin O at first under the effect of RNA primer in two zones
H, when heavy chain be synthesized to whole mitochondrial DNA 2/3rds the time, part parent's heavy chain of cementing out arrives light chain replication origin (O
L), this site is arranged in five tRNA gene clusters; After the light chain replication origin is displaced, O
LBe folded into hairpin structure, archaeal dna polymerase is the synthetic of the initial new light chain of template with the heavy chain that displaces; Mitochondrial DNA carries out asynchronous two-way synthetic subsequently.
Mitochondrial DNA is matrilinear inheritance, and mitochondrial DNA can reach 100,000 copies in the single egg mother cell, yet mitochondrial DNA only contains 100～1500 copies in sperm.Mitochondria in the fertilization process sperm enters egg mother cell, but fades away at embryogenetic two-cell stage and four cell stage.Method by artificial insemination and single-semen injection is grown in the individuality that forms, from the mitochondrial DNA of paternal line also detect less than.
Mitochondria self lacks effective DNA repair system, simultaneously the also protein that defencive function is arranged resemble the histone not; And mitochondrial DNA also is exposed to by strong mutagens---the oxygen radical that produces in the OXPHOS process.Unusual metabolism may also can accumulate the mitochondrial DNA of sudden change in the mitochondria in addition.These relatively unique character have caused the polymorphic accumulating rate of mitochondrial DNA than fast 10～17 times of nuclear DNA.Comparing the mutation rate of the noncoding region of mitochondrial DNA with the code area wants high 5-10 doubly.
The mitochondrial DNA noncoding region is the high catastrophe characteristics in D-loop district just, and the characteristic that the height of mitochondrial DNA copies in unicellular, make it compare the huge advantage that exists with nuclear DNA in sample is differentiated: (1) has relative high detection sensitivity to the sample of highly degraded, (2) can detect the sample of trace, (3) high mutation rate makes it have to be had between higher species or the specificity in the species than nuclear DNA.
Summary of the invention
An object of the present invention is to provide a kind of method of differentiating the ordinary milk that whether mixes in the yak dairy produce.
The method of differentiating the ordinary milk that whether mixes in the yak dairy produce provided by the present invention, may further comprise the steps: be template with the DNA that extracts from yak dairy produce to be measured 1), utilizing nucleotide sequence is respectively that a pair of primer of sequence 1 in the sequence table and sequence 2 carries out the PCR reaction;
2) detect in the pcr amplification product whether the DNA band is arranged,, be doped with ordinary milk in the yak dairy produce then to be measured as the DNA band is arranged in the pcr amplification product.
The size of described DNA band can be 445bp (to all common cattle breeds or individualities, this DNA stripe size all is 445bp).
Described yak can be all yaks (Bos grunniens) kind.Though all can there be the difference of indivedual bases in the D-loop district between different yak kinds or the individuality, to not influence of pcr amplification of the present invention, carry out pcr amplification with a pair of primer of sequence in the above-mentioned sequence table 1 and sequence 2, all can not get the DNA band.
Described common ox can be the cattle breeds of all Bos taurus kinds.Though all can there be the difference of indivedual bases in the D-loop district between different common cattle breeds or the individuality, but to not influence of pcr amplification of the present invention, a pair of primer with sequence in the above-mentioned sequence table 1 and sequence 2 carries out pcr amplification, all obtains the DNA band of 445bp.
Also comprise in the said method yak dairy produce to be measured is carried out pretreated step; Described pretreated method is for to carry out the dialysis that molecular cut off is 8000-12000Da with yak dairy produce to be measured, add Tris-Cl in the yak dairy produce after dialysis then, EDTA, NaCl and Proteinase K, the final concentration that makes Tris-Cl is that the final concentration of 10mmol/L, EDTA is that the final concentration of 5mmol/L, NaCl is that the final concentration of 50mmol/L, Proteinase K is 200ug/ml, 50 ℃ of reactions 2 hours.
Second purpose of the present invention provides a kind of whether mix a pair of primer of ordinary milk of yak dairy produce that is used for differentiating.
Provided by the present inventionly be used for differentiating whether the mix a pair of primer of ordinary milk of yak dairy produce, its nucleotide sequence is respectively sequence 1 and the sequence 2 in the sequence table.
Another purpose of the present invention provides a kind of whether mix kit of ordinary milk of yak dairy produce that is used for differentiating.
Provided by the present inventionly be used for differentiating whether the mix kit of ordinary milk of yak dairy produce, it contains nucleotide sequence is respectively the sequence 1 in the sequence table and a pair of primer of sequence 2.
Described kit also comprises the PCR reaction reagent.
The present invention can take some body cells (average 3.9 * 10 according to emulsion out of thereupon in extrusion
4Individual/milliliter), from dairy produce, extract the DNA of trace, and there is bigger difference in the D-Loop district of mitochondrial DNA between yak and common ox, designs the special primer of common milk cow according to the difference between them, identifies mingling the yak dairy produce by PCR method.
The method of ordinary milk of whether mixing in the discriminating of the present invention yak dairy produce (only needs 6 hours) fast, accurate, sensitive.
Description of drawings
Fig. 1 is the difference between Chinese yak breast and colostrum nutritional labeling average content and the common milk cow.
Fig. 2 is an existing difference between common ox D-loop (NC_006853) and the yak D-loop (NC_006380).
Fig. 3 is the yak of primer Milk-id-F and Milk-id-R and the amplification of common ox mixed emulsion.
Fig. 4 is the yak of primer Milk-id-F and Milk-id-R and the amplification that common ox mixes milk powder.
The specific embodiment
The present invention has analyzed the difference between yak mitochondrial DNA D-loop region sequence (NC_006380, sequence 4 in the sequence table) and the common ox mitochondrial DNA D-loop region sequence (NC_006853, sequence 3 in the sequence table).The result as shown in Figure 2, Yak is the sequence in yak mitochondrial DNA D-loop district in the sequence, Cattle is the sequence in common milk cow mitochondrial DNA D-loop district, Majority is the conserved sequence between them.The forward and reverse primer of box indicating position.Common ox among Fig. 2 is meant He Sitanniu, and yak is meant the Qinghai yak.Though all can there be the difference of indivedual bases in the D-loop district between different yak kinds or the individuality, not influence concerning pcr amplification of the present invention is carried out pcr amplification with a pair of primer of sequence in the above-mentioned sequence table 1 and sequence 2, all can not get the DNA band; Though all can there be the difference of indivedual bases in the D-loop district between different common cattle breeds or the individuality, but not influence concerning pcr amplification of the present invention, a pair of primer with sequence in the above-mentioned sequence table 1 and sequence 2 carries out pcr amplification, all obtains the DNA band of 445bp.
Difference according between yak and the common ox mitochondrial DNA D-loop district, designed the different primer of common Newt:
Milk-id-F:5 '-GCCCATACACAGACCAC-3 ' (sequence 1)
Milk-id-R:5 '-GTGAGCATGGGCTGATT-3 ' (sequence 2).
Experimental technique among the following embodiment if no special instructions, is conventional method.
Experiment dairy products: Qinghai yak milk and milk powder that He Sitanniu ordinary milk and milk powder that market is buied, market are buied.
Yak milk and ordinary milk mix by 1: 1,3: 1,6: 1,10: 1,20: 1 volume ratio respectively, identify the ordinary milk composition in the yak milk.
Yak milk powder and ordinary milk powder by 1: 1,3: 1,6: 1,10: 1,20: 1 mass ratio mixing, are identified the ordinary milk meal component in the yak milk powder respectively.
1, the extraction of DNA in the milk
(1) gets milk 500 μ l, put into the bag filter that molecular cut off is 8000-12000Da (dalton).
(2) bag filter is put into ready 500ml distilled water, dialysed 1 hour.
(3) the milk sample in the bag filter is moved in the EP pipe of 1ml.
(4) add Tris-Cl, EDTA, NaCl and Proteinase K, the final concentration that makes Tris-Cl are that the final concentration of 10mmol/L, EDTA is that the final concentration of 5mmol/L, NaCl is that the final concentration of 50mmol/L, Proteinase K is 200ug/ml, 50 ℃ of reactions 3 hours.
(5) add the saturated phenol of isopyknic Tris-, extracting 10min, the centrifugal 10min of 11000g.
(6) get supernatant repeating step (5).
(7) get the phenol chloroform extracting 10min that supernatant adds isopyknic 1: 1 (volume ratio), the centrifugal 10min of 11000g.
(8) get the absolute ethyl alcohol that supernatant adds two volumes, the centrifugal 10min of 11000g.
(9) abandon supernatant, twice of the ethanol rinsing of 70% (volumn concentration).After 37 ℃ of oven dry, dissolve in the distilled water of 5ul.
Identical method is extracted 10 parts of same samples, and the DNA that extracts is incorporated in the individual system.
2, the extraction of DNA in the milk powder
(1) get milk powder 100 μ g, dissolve in the 600 μ l distilled waters, the immigration molecular cut off is in 8000-12000Da (dalton's) the bag filter.
(2) bag filter is put into ready 500ml distilled water, dialysed 2 hours.
(3) change the 500ml distilled water, continue dialysis 3 hours.
(4) the milk sample in the bag filter is moved in the EP pipe of 1ml every pipe 500ul milk sample.
(5) add Tris-Cl, EDTA, NaCl and Proteinase K, the final concentration that makes Tris-Cl are that the final concentration of 10mmol/L, EDTA is that the final concentration of 5mmol/L, NaCl is that the final concentration of 50mmol/L, Proteinase K is 200ug/ml, 50 ℃ of reactions 3 hours.
(6) add the saturated phenol of isopyknic Tris-, extracting 10min, the centrifugal 10min of 11000g.
(7) get supernatant repeating step (6).
(8) get the phenol chloroform extracting 10min that supernatant adds isopyknic 1: 1 (volume ratio), the centrifugal 10min of 11000g.
(9) get the absolute ethyl alcohol that supernatant adds two volumes, the centrifugal 10min of 11000g.
(10) abandon supernatant, twice of the ethanol rinsing of 70% (volumn concentration).After 37 ℃ of oven dry, dissolve in the distilled water of 5ul.
Identical method is extracted 10 parts of same samples, and the DNA that extracts is incorporated in the individual system.
3, PCR reaction and detection
The DNA that extracts with step 1 and step 2 is a template respectively, utilizes Milk-id-F:
5 '-GCCCATACACAGACCAC-3 ' (sequence 1) and Milk-id-R:5 '-GTGAGCATGGGCTGATT-3 ' (sequence 2) carry out the PCR reaction.Wherein, the PCR reaction system is as follows:
10 times amplification buffer 2.5ul
4 kinds of dNTP mixtures, (each 42.5mmol/L) 2ul
Milk-id-F and Milk-id-R (each 10pmol/ul) 1ul
Template DNA 5ul
Taq archaeal dna polymerase (3u/ul) 1ul
ddH
2O 13.5ul
Cumulative volume 25ul
Reaction condition:
94 ℃ of pre-sex change 5 minutes; Cycling condition (72 ℃ were extended 30 seconds for 94 ℃ of sex change 30 seconds, 65 ℃ of annealing 30 seconds), 35 circulations; 72 ℃ were extended 7 minutes; 4 ℃ of preservations.Product detects with 2% Ago-Gel, and amplified production is checked order.
The testing result of yak milk does not obtain the DNA band in the yak milk in contrast as shown in Figure 3, and yak milk and ordinary milk all obtain the DNA band of 445bp respectively in the sample by 1: 1,3: 1,6: 1,10: 1,20: 1 volume ratio mixing.And the sample of not sneaking into common milk does not have amplified band to produce, and this result shows that primer Milk-id-F and Milk-id-R are specific to common ox, can be used for differentiating the ordinary milk that whether mixes in the yak dairy produce.Among Fig. 3, M is the 100bp molecular weight marker, and CON is that pcr template is the contrast of yak milk, and 1-5 is respectively yak milk and ordinary milk respectively by 20: 1,10: 1, and 5: 1, the composite sample PCR result of the volume ratio of 3: 1 and 1: 1.
The testing result of yak milk powder as shown in Figure 4, do not obtain the DNA band in the yak milk powder in contrast, all obtain the DNA band of 445bp in the sample that yak milk powder and ordinary milk powder mix by 1: 1,3: 1,6: 1,10: 1,20: 1 mass ratio respectively.And the sample of not sneaking into ordinary powdered milk does not have amplified band, illustrates that primer Milk-id-F and Milk-id-R are specific to common ox, can be used for differentiating the ordinary milk that whether mixes in the yak dairy produce.Among Fig. 4, M is the pBR322 molecular weight marker, and CON is that pcr template is the contrast of yak milk powder, and 1-5 is respectively yak milk powder and ordinary milk powder respectively by 20: 1,10: 1, and 5: 1, the composite sample PCR result of the mass ratio of 3: 1 and 1: 1.
Sequence table
<160>4
<210>1
<211>17
<212>DNA
＜213〉artificial sequence
<220>
<223>
<400>1
gcccatacac?agaccac 17
<210>2
<211>17
<212>DNA
＜213〉artificial sequence
<220>
<223>
<400>2
gtgagcatgg?gctgatt 17
<210>3
<211>910
<212>DNA
＜213〉ox (Bos taurus)
<400>3
aacactatta?atatagttcc?ataaatacaa?agagccttat?cagtattaaa?tttatcaaaa 60
atcccaataa?ctcaacacag?aatttgcacc?ctaaccaaat?attacaaaca?ccactagcta 120
acataacacg?cccatacaca?gaccacagaa?tgaattacct?acgcaagggg?taatgtacat 180
aacattaatg?taataaagac?ataatatgta?tatagtacat?taaattatat?gccccatgca 240
tataagcaag?cacatgacct?ctatagcagt?acataataca?tataattatt?gactgtacat 300
aatacattat?gtcaaattca?ttcttgatag?tatatctatt?atatattcct?taccattaga 360
tcacgagctt?aattaccatg?ccgcgtgaaa?ccagcaaccc?gctaggcagg?gatccctctt 420
ctcgctccgg?gcccataaac?cgtgggggtc?gctatccaat?gaattttacc?aggcatctgg 480
ttctttcttc?agggccatct?catctaaaac?ggtccattct?ttcctcttaa?ataagacatc 540
tcgatggact?aatggctaat?cagcccatgc?tcacacataa?ctgtgctgtc?atacatttgg 600
tattttttta?ttttggggga?tgcttggact?cagctatggc?cgtcaaaggc?cctgacccgg 660
agcatctatt?gtagctggac?ttaactgcat?cttgagcacc?agcataatga?taagcatgga 720
cattacagtc?aatggtcaca?ggacataaat?tatattatat?atcccccctt?cataaaaatt 780
tcccccttaa?atatctacca?ccacttttaa?cagacttttc?cctagatact?tatttaaatt 840
tttcacgctt?tcaatactca?atttagcact?ccaaacaaag?tcaatatata?aacgcaggcc 900
cccccccccc 910
<210>4
<211>894
<212>DNA
＜213〉yak (Bos grunniens)
<400>4
aacgctatta?atatagttcc?ataaatgtaa?agagcctcac?cagtattaaa?tttactaaaa 60
attccaataa?ctcaacacaa?actttgtact?ctaaccaaat?attacaaaca?ccactagcta 120
acaacacaca?tccccaaaaa?tgcattgtcc?aaacggggga?tacgtacata?atattaatgt 180
aataaagaca?tattatgtat?atagtacatt?aaattatatg?ccccatgcat?ataagcaagt 240
acataattcc?tattgatagt?acatagtaca?tgaaattatt?aatcgtacat?aacacattat 300
gtcaaaccta?ctcctaacaa?catgtatgcc?ccctccacta?gatcacgagc?ttaactacca 360
tgccgggtga?aaccagcaac?ccgctaggcg?ggggactcct?cttcttggct?cccgggccca 420
tgaactgtgg?gggtcgctat?ttaatgaact?ttatcagaca?tctggttctt?tcttcagggc 480
catctcacct?aaaaccgtcc?actctttcct?cttaaataag?acatctcgat?ggactaatgg 540
ctaatcagcc?catgctcaca?cataactgtg?ctgtcataca?tttggtattt?ttttattttg 600
ggggatgctt?ggactcagct?atggccgtca?aaggccccga?cccggagcat?ctattgtagc 660
tggacttaac?tgcatcttga?gcaccagcat?aatgataggc?atgcacatat?agtcaatggt 720
cacaggacat?agattatatt?atatatcccc?ccctccataa?aaattccccc?ttaaatattt 780
accactactt?ttaacagatt?tttccctagt?tgcttattta?aaatttccac?actttcaata 840
ctcaaattag?cactccatat?aaagtcaata?tataaacgca?gggccccccc?cccc 894
Claims (9)
1, a kind of method of differentiating the ordinary milk that whether mixes in the yak dairy produce, may further comprise the steps: be template with the DNA that extracts from yak dairy produce to be measured 1), utilizing nucleotide sequence is respectively that a pair of primer of sequence 1 in the sequence table and sequence 2 carries out the PCR reaction;
2) detect in the pcr amplification product whether the DNA band is arranged,, be doped with ordinary milk in the yak dairy produce then to be measured as the DNA band is arranged in the pcr amplification product.
2, method according to claim 1 is characterized in that: the size of described DNA band is 445bp.
3, method according to claim 1 and 2 is characterized in that: described yak is all yak kinds of Bos grunniens kind.
4, method according to claim 1 and 2 is characterized in that: described common ox is all cattle breeds of Bos taurus kind.
5, method according to claim 1 is characterized in that: described yak is the Qinghai yak, and described common ox is He Sitanniu.
6, according to arbitrary described method in the claim 1 to 5, it is characterized in that: also comprise in the described method yak dairy produce to be measured is carried out pretreated step; Described pretreated method is for to carry out the dialysis that molecular cut off is 8000-12000Da with yak dairy produce to be measured, add Tris-Cl in the yak dairy produce after dialysis then, EDTA, NaCl and Proteinase K, the final concentration that makes Tris-Cl is that the final concentration of 10mmol/L, EDTA is that the final concentration of 5mmol/L, NaCl is that the final concentration of 50m mol/L, Proteinase K is 200ug/ml, 50 ℃ of reactions 2 hours.
7, be used for differentiating whether the mix a pair of primer of ordinary milk of yak dairy produce, its nucleotide sequence is respectively sequence 1 and the sequence 2 in the sequence table.
8, be used for differentiating whether the mix kit of ordinary milk of yak dairy produce, it contains nucleotide sequence is respectively the sequence 1 in the sequence table and a pair of primer of sequence 2.
9, kit according to claim 8 is characterized in that: described kit also comprises the PCR reaction reagent.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2006101143296A CN100544603C (en) | 2006-11-06 | 2006-11-06 | Differentiate the method and the primer special thereof of the ordinary milk that whether mixes in the yak dairy produce |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2006101143296A CN100544603C (en) | 2006-11-06 | 2006-11-06 | Differentiate the method and the primer special thereof of the ordinary milk that whether mixes in the yak dairy produce |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1965664A true CN1965664A (en) | 2007-05-23 |
| CN100544603C CN100544603C (en) | 2009-09-30 |
Family
ID=38074781
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2006101143296A Expired - Fee Related CN100544603C (en) | 2006-11-06 | 2006-11-06 | Differentiate the method and the primer special thereof of the ordinary milk that whether mixes in the yak dairy produce |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100544603C (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110055336A (en) * | 2019-02-26 | 2019-07-26 | 中国农业科学院农业质量标准与检测技术研究所 | A kind of kit whether quick identification yak milk is adulterated |
| CN117051115A (en) * | 2022-09-20 | 2023-11-14 | 哈尔滨海关技术中心 | Primer group for identifying species and germ line of incoming and outgoing cattle and pigs and application of primer group |
-
2006
- 2006-11-06 CN CNB2006101143296A patent/CN100544603C/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110055336A (en) * | 2019-02-26 | 2019-07-26 | 中国农业科学院农业质量标准与检测技术研究所 | A kind of kit whether quick identification yak milk is adulterated |
| CN110055336B (en) * | 2019-02-26 | 2022-04-05 | 中国农业科学院农业质量标准与检测技术研究所 | Kit for rapidly identifying whether yak milk is adulterated or not |
| CN117051115A (en) * | 2022-09-20 | 2023-11-14 | 哈尔滨海关技术中心 | Primer group for identifying species and germ line of incoming and outgoing cattle and pigs and application of primer group |
| CN117051115B (en) * | 2022-09-20 | 2024-04-30 | 哈尔滨海关技术中心 | Primer group for identifying species and germ line of incoming and outgoing cattle and pigs and application of primer group |
Also Published As
| Publication number | Publication date |
|---|---|
| CN100544603C (en) | 2009-09-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Chen et al. | Effects of fishmeal replacement with cottonseed protein concentrate on growth, digestive proteinase, intestinal morphology and microflora in pearl gentian grouper (♀ Epinephelus fuscoguttatus×♂ Epinephelus lanceolatu) | |
| CN104250646B (en) | A kind of molecular labeling relevant to pig feed transformation efficiency proterties and detection method and application | |
| Matsui et al. | Microbial diversity in ostrich ceca as revealed by 16S ribosomal RNA gene clone library and detection of novel Fibrobacter species | |
| Hansen et al. | The inclusion of plant protein in cod diets, its effects on macronutrient digestibility, gut and liver histology and heat shock protein transcription | |
| CN108103211B (en) | Microsatellite primer and identification method for tridacna elongata and tridacna nova | |
| CN103667279B (en) | The molecule marker of pig average daily gain genes involved Resistin and application thereof | |
| Sarropoulou et al. | Comparative genomics in teleost species: knowledge transfer by linking the genomes of model and non-model fish species | |
| CN104372086B (en) | A kind of primer and method of fast detecting fugu obscurus juvenile fish gender differences single base mutation | |
| CN110777210A (en) | Siniperca chuatsi male molecular marker primer and application thereof | |
| Meyer et al. | DNA fingerprinting of pearls to determine their origins | |
| CN1789974A (en) | Molecular biology method for quick identification of fish | |
| Johny et al. | A primer on metagenomics and next‐generation sequencing in fish gut microbiome research | |
| CN1158390C (en) | Prolactin receptor gene as genetic marker for increased litter size in pigs | |
| Wan et al. | Dynamic expression of HSP90B1 mRNA in the hypothalamus of two Chinese chicken breeds under heat stress and association analysis with a SNP in Huainan chickens | |
| Foysal et al. | Sources of protein diet differentially stimulate the gut and water microbiota under freshwater crayfish, marron (Cherax cainii, Austin 2002) culture | |
| CN1965664A (en) | Method for determining if common cow milk has been doped in yak milk product and primer dedicated therefor | |
| Tian et al. | Distribution of microbiota across different intestinal tract segments of a stranded dwarf minke whale, Balaenoptera acutorostrata | |
| CN101029340A (en) | Method for screening high-reproduction goat and its special primer | |
| Gao et al. | Ontogeny of the stomach in yellow catfish (Pelteobagrus fulvidraco): detection and quantifictation of pepsinogen and H+/K+‐ATPase gene expression | |
| Mishra et al. | Analysis of genetic diversity in Berari goat population of Maharashtra state | |
| CN103421769A (en) | SNP molecular marker related to pig carcass trait and application thereof | |
| CN103305506B (en) | SNP (single nucleotide polymorphism) molecular mark related to domesticating property of mandarin fish | |
| Yao et al. | Polymorphisms of LAP3 gene and their association with the growth traits in the razor clam Sinonovacula constricta | |
| CN105603097B (en) | Microsatellite marker primer for identifying pinctada fucata microsatellite families as well as identification method and application | |
| Tang et al. | Effects of sodium butyrate stimulation on immune‐related mRNA‐miRNA network in intestine of grass carp |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20090930 Termination date: 20121106 |