CN107881247A - The related molecular labeling of Red Steppe fat content and its acquisition methods and application - Google Patents
The related molecular labeling of Red Steppe fat content and its acquisition methods and application Download PDFInfo
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Abstract
The present invention provides the related molecular labeling of Red Steppe fat content and its acquisition methods and application, belongs to biotechnology and Agricultural production field.The acquisition methods of the related molecular labeling of Red Steppe fat content provided by the invention, pass through the experimental method of simple and fast, the related molecular labeling of accurate Red Steppe fat content can be obtained, the related molecular labeling of the Red Steppe fat content of acquisition is applied in the identification of Red Steppe meat fat content matter, it preferably can distinguish and filter out more high-quality meat, therefore the related molecular labeling of Red Steppe fat content can preferably instruct the production of farming and animal husbandry especially animal husbandry, have larger use value and social value.
Description
Technical field
The present invention relates to biotechnology and Agricultural production field, related in particular to Red Steppe fat content
Molecular labeling and its acquisition methods and application.
Background technology
The meat new lines of China Grassland Red Cattle are passed through by beef cattle research team of Pasturage Science School of Jilin Agricultural Sciences Academy
After seed selection for many years into the meat new lines seed selection success of China Grassland Red Cattle, new work is filled with to the development of China's beef cattle industries
Power, research of the domestic and international Genetic Improvement of Beef Cattle expert to slaughter trait is extensive not as growth traits research, because
These slaughter trait data obtain relative difficult, just bigger particularly with degree-of-difficulty factor for big livestock animals, so as to certain
The research of slaughter trait genetic marker is limited in degree " but the today improved constantly in people's living standard, people are to ox
The requirement of meat also more and more higher, this requires Genetic Improvement of Beef Cattle expert to be continuing effort to cultivate more preferable beef breed to expire
" in view of considerations above, domestic and international Genetic Improvement of Beef Cattle expert is increased to beef cattle the demand of sufficient people using marker assisted selection technology
The research dynamics of slaughter trait genetic marker, so as to directly to carry out Seedling selection using genotype, shorten the generation inteval, add
Fast genetic progress is laid a good foundation.
So far, the polymorphism in the meat new lines colony of China Grassland Red Cattle to whole gene and its with butchering property
At home and abroad there is not been reported for the relation research of shape.
The content of the invention
The first object of the present invention is the acquisition methods for providing the related molecular labeling of Red Steppe fat content, and this is obtained
Experimental method of the method by simple border is taken, can quickly obtain the related molecular labeling of Red Steppe fat content.
The second object of the present invention is the molecular labeling for providing Red Steppe fat content correlation, the molecular labeling of acquisition
With preferable application value, can preferably instruct in farming and animal husbandry, especially livestock breed aquatics.
The second object of the present invention is to provide the related molecular labeling of the Red Steppe fat content stated in Red Steppe
Application in the identification of meat fat content.
In order to realize the above-mentioned purpose of the present invention, using following technical scheme：
The acquisition methods of the related molecular labeling of Red Steppe fat content, comprise the following steps：
The genomic DNA of Red Steppe is extracted, and with the first enzyme liquid digestion process, obtains purified genomic dna；
550-800ng purified genomic dna denaturation treatment is taken, obtains being denatured genomic DNA；
Denaturation genomic DNA is expanded, and obtains genome amplification product；Genome amplification product through the second enzyme liquid at
Reason, obtains sample DNA fragment；
React, and fluorescent scanning and analysis, obtain and Red Steppe with DNA chip after sample DNA fragment Single base extension
The related molecular labeling of fat content.
The related molecular labeling of Red Steppe fat content, the related molecular labeling of Red Steppe fat content pass through as above
The acquisition methods of the molecular labeling for the Red Steppe fat content correlation stated obtain
Application of the related molecular labeling of above-mentioned Red Steppe fat content in the identification of Red Steppe meat fat content.
Compared with prior art, beneficial effects of the present invention are：Red Steppe fat content correlation provided by the invention
The acquisition methods of molecular labeling, by the experimental method of simple and fast, accurate Red Steppe fat content correlation can be obtained
Molecular labeling, the related molecular labeling of the Red Steppe fat content of acquisition is applied to the identification of Red Steppe meat fat content matter
In, more high-quality meat preferably can be distinguished and filter out, therefore the molecular labeling of Red Steppe fat content correlation can be more
The good production for instructing farming and animal husbandry especially animal husbandry, has larger use value and social value.
Embodiment
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 present invention.It is unreceipted specific in embodiment
Condition person, the condition suggested according to normal condition or manufacturer are carried out.Agents useful for same or the unreceipted production firm person of instrument, it is
The conventional products that can be obtained by commercially available purchase.
The related molecular labeling of Red Steppe fat content to the embodiment of the present invention and its acquisition methods and application below
It is specifically described.
The acquisition methods of the related molecular labeling of Red Steppe fat content, comprise the following steps：
The genomic DNA of Red Steppe is extracted, and with the first enzyme liquid digestion process, obtains purified genomic dna；
The 500-700ng purified genomic dna denaturation treatment is taken, obtains being denatured genomic DNA；
Denaturation genomic DNA is expanded, and obtains genome amplification product；Genome amplification product through the second enzyme liquid at
Reason, obtains sample DNA fragment；
React, and fluorescent scanning and analysis, obtain and Red Steppe with DNA chip after sample DNA fragment Single base extension
The related molecular labeling of fat content.
Further, in presently preferred embodiments of the present invention, the A260/A280 values of genomic DNA are 1.8-2.1.
Further, in presently preferred embodiments of the present invention, the concentration of genomic DNA is 500ng/ μ L-2000ng/ μ L.
If A260/A280 values, which are less than in 1.8 expression samples, has the problem of Substances Pollutions such as protein, phenol, if
The problem of A260/A280 values show to there may be RNA pollutions more than 2.1.
Further, in presently preferred embodiments of the present invention, the concentration of genomic DNA is 650ng/ μ L-2500ng/ μ L.
Different collagenase treatments is selected according to the A260/A280 values of genomic DNA, can if A260/A280 values are larger
To be handled from RNase as the first enzyme liquid, if A260/A280 values are small, protease can be selected as the first enzyme liquid；Certainly
It can also be handled successively with RNase and protease simultaneously.
Further, in presently preferred embodiments of the present invention, the first enzyme liquid is at least one of RNase or protease.
Further, in presently preferred embodiments of the present invention, the denaturant of denaturation treatment is NaOH solution.
Further, in presently preferred embodiments of the present invention, the second enzyme liquid includes restriction endonuclease.
Genome is cut into by suitable short-movie section by restriction endonuclease, short-movie section just can be good at and chip
Upper sequence hybridization reaction.
Further, in presently preferred embodiments of the present invention, after restriction endonuclease processing genome amplification product also
Including precipitating and being resuspended, the reaction reagent of precipitation is ethanol solution.
Further, in presently preferred embodiments of the present invention, the related molecular labeling of Red Steppe fat content includes thick fat
Fat content molecular labeling, meat weight molecular labeling and carcass weight molecular labeling；The crude fat content molecular labeling includes
ELAVL2 genetic mutation sites and ELAVL2 genetic mutation sites, the meat weight molecular labeling include LOC104970972 genes
Variant sites, RAP2B genetic mutation sites, MBNL1 genetic mutation sites and CCDC30 genetic mutation sites, the carcass weight point
Son mark include LTA4H genetic mutation sites, C7H5orf63 genetic mutation sites, LOC101907891 genetic mutation sites,
KCNS3 genetic mutation sites, LOC786289 genetic mutation sites and FER1L6 genetic mutation sites.
The LTA4H genetic mutation sites of carcass weight molecular labeling are loc60711494, C7H5orf63 genetic mutation site
It is that loc10665717, KCNS3 genetic mutation site are for loc28343212, LOC101907891 genetic mutation site
Loc80905529, LOC786289 genetic mutation site are that loc60324725 and FER1L6 genetic mutation sites are
Loc17551045, and MED30 genetic mutation sites are loc49023940 and loc49032157.
The related molecular labeling of Red Steppe fat content, the related molecular labeling of Red Steppe fat content pass through as above
The acquisition methods of the related molecular labeling of Fang Shu Red Steppe fat content obtain.
Application of the related molecular labeling of above-mentioned Red Steppe fat content in the identification of Red Steppe meat fat content.
The feature and performance of the present invention are described in further detail with reference to embodiments.
Embodiment 1
The present embodiment provides a kind of acquisition methods of the related molecular labeling of Red Steppe fat content, and specific steps are such as
Under：
Sampling：The meat new lines of Red Steppe 48 are randomly selected, jugular vein blood collection, jugular vein blood collection is put into added with EDTA
In the centrifuge tube of anti-coagulants, then shake up and be put into ice chest rapidly, -80 DEG C of preservations are stand-by；
Red Steppe carcass weight, meat weight, carry out on-site measurement and to dressing percentage, the acquisition of the data of neat percentage.
The Red Steppe of selection is slaughtered, according to People's Republic of China (PRC) agricultural industry criteria NY/T676-2003《Beef matter
Amount classification》To carcass weight, meat weight, progress on-site measurement and dressing percentage, neat percentage are calculated；
1.1 carcass weight：Refer to decaptitating, skin, tail, hoof, reproductive organs and peripheral adipose, internal organ (retaining kidney and peripheral adipose)
Weight.Meat weight：Trunk rejects the total weight after bone, including kidney and peripheral adipose.Dressing percentage (%)=(carcass weight/
Slaughter weight) × 100%, neat percentage (%)=(meat weight/Slaughter weight) × 100%.Measure meat weight simultaneously.
1.2 percentage of water loss：2h takes longissimus dorsi muscle 1.0cm chop samples after government official, is cut with diameter 2.532cm rounded sample devices
Lower area 5cm2, thick 1cm circular meat sample is standby after weighing.After soil permission dilatometer steel loop first is forced into maximum three times
It is decompressed to amesdial back to zero.Then circular meat sample is clipped among two layers of gauze, respectively pads 18 layers of qualitative Medium speed filter paper of Xinhua up and down,
It is sandwiched in again between two layers of duroplasts, being placed in soil allows to be forced into 35kg holdings 5min on expansion compressometer platform.After removing pressure
Immediately meat sample is peeled from gauze to weigh.
Percentage of water loss={ (meat sample weight before pressure-meat sample weight before pressure)/meat sample weight before pressure } × 100%
The measure of 1.3 conventional nutrients：Take longissimus dorsi muscle at 12-13 thoracic vertebraes, after rubbing, conventionally carry out moisture,
Intramuscular fat, coarse ash, the measure of crude protein.Measurement result is represented with the content of conventional nutrient in fresh meat sample.
It is 1.4 yellowish pink：By 5 fraction yellowish pink scale figure visual scores.Two/allow to set 0.5 score value.
1.5 marbling：According to《Chinese beef cattle classification standard》Middle marbling grade collection of illustrative plates (wherein marble grain
What grade figure provided is the minimum standard of decorative pattern in every grade) determine marbling grade at eye muscle cross section.Marbling
Grade is divided into 5 grades:1 grade, 2 grades, 3 grades, 4 grades, 5 grades.Marbling is enriched for 5 grades；Relatively enrich for 4 grades, generally 3
Level, a small amount of is 2 grades, almost without for 1 grade.
1.6 Meat Tenderness, are represented with shearing force, take 13-16 thoracic vertebraes portion to intercept thickness in vertical direction by with muscle fibre
About 2.5cm longissimus dorsi muscle, the fat of sample meat surface attachment is removed, is fitted into plastic film bag and bandages, be placed on 15
Carry out cadaveric rigidity pre-treatment within 24 hours under the conditions of DEG C -16 DEG C.Then meat sample is placed in refrigerator cold-storage layer, 24 is cured under the conditions of 4 DEG C
Hour.The meat sample that curing is completed is taken out, is placed 1 hour at room temperature.Then, plastic film packaging bag is opened, is inserted with thermometer
Enter muscle central part, then bandage meat sample, keep sack upward, be put into 80 DEG C of thermostat water baths, continuous heating after capping, directly
Untill muscle central temperature reaches 70 DEG C.Take out meat sample, placement make at room temperature muscle be cooled to 20 DEG C it is standby.Use again
The rounded sample device of 1.27cm diameters drills through cedductor along muscle fibre direction and (samples as much as possible, while it is noted that avoid muscle
Post), the shear force value of each cedductor is then determined with C-LM3 types digital display muscle tenderness instrument (Northeast Agricultural University's engineering science),
Each sample is surveyed 10--15 times, is chosen the 6-8 value that shearing force is close and is taken its average value, is to take cedductor shear force value.
Unit is represented with newton (N) or kilogram (kg).
The acquisition methods of the related molecular labeling of Red Steppe fat content, are comprised the following steps that：
2.1 extract genomic DNA from freezing with kit in blood sample, and detect the A260/A280 values of genomic DNA；
2.2 concentration for measuring genomic DNA are 500ng/ μ L, and first handle genomic DNA with RNase, then use protease
Genomic DNA solution is handled, obtains purified genomic dna, A260/A280 is 1.8 for control；
2.3 take 700ng purified genomic dna sample, and denaturation treatment is carried out using NaOH solution, obtain being denatured genome
DNA；
2.4 pairs of purified genomic dna samples expand, and obtain genome amplification product；
2.5 use KpnI and BamHI to obtain sample DNA fragment as the random digestion genome amplification product of the second enzyme liquid；
2.6 pairs of sample DNA fragments carry out single base extensions, carry out Two Colour Fluorescence dye marker, then with red ox DNA
Chip Bovine High Density chip carry out hybridization reaction；
After 2.7 cleaning chips, the fluorescence data of chip is scanned；
2.8 are obtained by analysis, the related molecular labeling of Red Steppe fat content include crude fat content molecular labeling,
Carcass weight molecular labeling and meat weight molecular labeling.
Embodiment 2
The present embodiment provides the present embodiment and provides a kind of obtaining for Red Steppe fat content related molecular labeling, specifically
Step is as follows：
Sampling：The meat new lines of Red Steppe 48 are randomly selected, jugular vein blood collection, jugular vein blood collection is put into added with EDTA
In the centrifuge tube of anti-coagulants, then shake up and be put into ice chest rapidly, -80 DEG C of preservations are stand-by；
Red Steppe carcass weight, meat weight, carry out on-site measurement and to dressing percentage, the acquisition of the data of neat percentage.
The Red Steppe of selection is slaughtered, according to People's Republic of China (PRC) agricultural industry criteria NY/T676-2003《Beef matter
Amount classification》To carcass weight, meat weight, progress on-site measurement and dressing percentage, neat percentage are calculated；
1.1 carcass weight：Refer to decaptitating, skin, tail, hoof, reproductive organs and peripheral adipose, internal organ (retaining kidney and peripheral adipose)
Weight.Meat weight：Trunk rejects the total weight after bone, including kidney and peripheral adipose.Dressing percentage (%)=(carcass weight/
Slaughter weight) × 100%, neat percentage (%)=(meat weight/Slaughter weight) × 100%.Determine meat weight simultaneously.
1.2 percentage of water loss：2h takes longissimus dorsi muscle 1.0cm chop samples after government official, is cut with diameter 2.532cm rounded sample devices
Lower area 5cm2, thick 1cm circular meat sample is standby after weighing.After soil permission dilatometer steel loop first is forced into maximum three times
It is decompressed to amesdial back to zero.Then circular meat sample is clipped among two layers of gauze, respectively pads 18 layers of qualitative Medium speed filter paper of Xinhua up and down,
It is sandwiched in again between two layers of duroplasts, being placed in soil allows to be forced into 35kg holdings 5min on expansion compressometer platform.After removing pressure
Immediately meat sample is peeled from gauze to weigh.
Percentage of water loss={ (meat sample weight before pressure-meat sample weight before pressure)/meat sample weight before pressure } × 100%
The measure of 1.3 conventional nutrients：Take longissimus dorsi muscle at 12-13 thoracic vertebraes, after rubbing, conventionally carry out moisture,
Intramuscular fat, coarse ash, the measure of crude protein.Measurement result is represented with the content of conventional nutrient in fresh meat sample.
It is 1.4 yellowish pink：By 5 fraction yellowish pink scale figure visual scores.Two/allow to set 0.5 score value.
1.5 marbling：According to《Chinese beef cattle classification standard》Middle marbling grade collection of illustrative plates (wherein marble grain
What grade figure provided is the minimum standard of decorative pattern in every grade) determine marbling grade at eye muscle cross section.Marbling
Grade is divided into 5 grades:1 grade, 2 grades, 3 grades, 4 grades, 5 grades.Marbling is enriched for 5 grades；Relatively enrich for 4 grades, generally 3
Level, a small amount of is 2 grades, almost without for 1 grade.
1.6 Meat Tenderness, are represented with shearing force, take 13-16 thoracic vertebraes portion to intercept thickness in vertical direction by with muscle fibre
About 2.5cm longissimus dorsi muscle, the fat of sample meat surface attachment is removed, is fitted into plastic film bag and bandages, be placed on 15
Carry out cadaveric rigidity pre-treatment within 24 hours under the conditions of DEG C -16 DEG C.Then meat sample is placed in refrigerator cold-storage layer, 24 is cured under the conditions of 4 DEG C
Hour.The meat sample that curing is completed is taken out, is placed 1 hour at room temperature.Then, plastic film packaging bag is opened, is inserted with thermometer
Enter muscle central part, then bandage meat sample, keep sack upward, be put into 80 DEG C of thermostat water baths, continuous heating after capping, directly
Untill muscle central temperature reaches 70 DEG C.Take out meat sample, placement make at room temperature muscle be cooled to 20 DEG C it is standby.Use again
The rounded sample device of 1.27cm diameters drills through cedductor along muscle fibre direction and (samples as much as possible, while it is noted that avoid muscle
Post), the shear force value of each cedductor is then determined with C-LM3 types digital display muscle tenderness instrument (Northeast Agricultural University's engineering science),
Each sample is surveyed 10-15 times, is chosen the 6-8 value that shearing force is close and is taken its average value, is to take cedductor shear force value.It is single
Position is represented with newton (N) or kilogram (kg).
The acquisition methods of the related molecular labeling of Red Steppe fat content, are comprised the following steps that：
2.1 extract genomic DNA from freezing with kit in blood sample, and detect the A260/A280 values of genomic DNA；
2.2 concentration for measuring genomic DNA are 2000ng/ μ L, and first handle genomic DNA with RNase, then use protease
Genomic DNA solution is handled, obtains purified genomic dna, A260/A280 is in the range of 2.1 or so for control；
2.3 take 500ng purified genomic dna sample, and denaturation treatment is carried out using NaOH solution, obtain being denatured genome
DNA；
2.4 pairs of purified genomic dna samples expand, and obtain genome amplification product；
2.5 use EcoRI and HindIII to obtain sample DNA piece as the random digestion genome amplification product of the second enzyme liquid
Section；
2.6 pairs of sample DNA fragments carry out single base extensions, carry out Two Colour Fluorescence dye marker, then with red ox DNA
Chip Bovine High Density chip carry out hybridization reaction；
After 2.7 cleaning chips, the fluorescence data of chip is scanned；
2.8 are obtained by analysis, and the related molecular labeling of Red Steppe fat content includes dressing percentage molecular labeling, trunk
Weight molecule mark, neat percentage molecular labeling and pure meat rate molecular labeling.
Embodiment 3
The present embodiment provides the acquisition that the present embodiment provides a kind of related molecular labeling of Red Steppe fat content, specifically
Step is as follows：
Sampling：The meat new lines of Red Steppe 48 are randomly selected, jugular vein blood collection, jugular vein blood collection is put into added with EDTA
In the centrifuge tube of anti-coagulants, then shake up and be put into ice chest rapidly, -80 DEG C of preservations are stand-by；
Red Steppe carcass weight, meat weight, carry out on-site measurement and to dressing percentage, the acquisition of the data of neat percentage.
The Red Steppe of selection is slaughtered, according to People's Republic of China (PRC) agricultural industry criteria NY/T676-2003《Beef matter
Amount classification》To carcass weight, meat weight, progress on-site measurement and dressing percentage, neat percentage are calculated；
1.1 carcass weight：Refer to decaptitating, skin, tail, hoof, reproductive organs and peripheral adipose, internal organ (retaining kidney and peripheral adipose)
Weight.Meat weight：Trunk rejects the total weight after bone, including kidney and peripheral adipose.Dressing percentage (%)=(carcass weight/
Slaughter weight) × 100%, neat percentage (%)=(meat weight/Slaughter weight) × 100%.Determine meat weight simultaneously.
1.2 percentage of water loss：2h takes longissimus dorsi muscle 1.0cm chop samples after government official, is cut with diameter 2.532cm rounded sample devices
Lower area 5cm2, thick 1cm circular meat sample is standby after weighing.After soil permission dilatometer steel loop first is forced into maximum three times
It is decompressed to amesdial back to zero.Then circular meat sample is clipped among two layers of gauze, respectively pads 18 layers of qualitative Medium speed filter paper of Xinhua up and down,
It is sandwiched in again between two layers of duroplasts, being placed in soil allows to be forced into 35kg holdings 5min on expansion compressometer platform.After removing pressure
Immediately meat sample is peeled from gauze to weigh.
Percentage of water loss={ (meat sample weight before pressure-meat sample weight before pressure)/meat sample weight before pressure } × 100%
The measure of 1.3 conventional nutrients：Take longissimus dorsi muscle at 12-13 thoracic vertebraes, after rubbing, conventionally carry out moisture,
Intramuscular fat, coarse ash, the measure of crude protein.Measurement result is represented with the content of conventional nutrient in fresh meat sample.
It is 1.4 yellowish pink：By 5 fraction yellowish pink scale figure visual scores.Two/allow to set 0.5 score value.
1.5 marbling：According to《Chinese beef cattle classification standard》Middle marbling grade collection of illustrative plates (wherein marble grain
What grade figure provided is the minimum standard of decorative pattern in every grade) determine marbling grade at eye muscle cross section.Marbling
Grade is divided into 5 grades:1 grade, 2 grades, 3 grades, 4 grades, 5 grades.Marbling is enriched for 5 grades；Relatively enrich for 4 grades, generally 3
Level, a small amount of is 2 grades, almost without for 1 grade.
1.6 Meat Tenderness, are represented with shearing force, take 13-16 thoracic vertebraes portion to intercept thickness in vertical direction by with muscle fibre
About 2.5cm longissimus dorsi muscle, the fat of sample meat surface attachment is removed, is fitted into plastic film bag and bandages, be placed on 15
Carry out cadaveric rigidity pre-treatment within 24 hours under the conditions of DEG C -16 DEG C.Then meat sample is placed in refrigerator cold-storage layer, 24 is cured under the conditions of 4 DEG C
Hour.The meat sample that curing is completed is taken out, is placed 1 hour at room temperature.Then, plastic film packaging bag is opened, is inserted with thermometer
Enter muscle central part, then bandage meat sample, keep sack upward, be put into 80 DEG C of thermostat water baths, continuous heating after capping, directly
Untill muscle central temperature reaches 70 DEG C.Take out meat sample, placement make at room temperature muscle be cooled to 20 DEG C it is standby.Use again
The rounded sample device of 1.27cm diameters drills through cedductor along muscle fibre direction and (samples as much as possible, while it is noted that avoid muscle
Post), the shear force value of each cedductor is then determined with C-LM3 types digital display muscle tenderness instrument (Northeast Agricultural University's engineering science),
Each sample is surveyed 10-15 times, is chosen the 6-8 value that shearing force is close and is taken its average value, is to take cedductor shear force value.It is single
Position is represented with newton (N) or kilogram (kg).
The acquisition methods of the related molecular labeling of Red Steppe fat content, are comprised the following steps that：
2.1 extract genomic DNA from freezing with kit in blood sample, and detect the A260/A280 values of genomic DNA；
2.2 concentration for measuring genomic DNA are 1500ng/ μ L, and first handle genomic DNA with RNase, then use protease
Genomic DNA solution is handled, obtains purified genomic dna, A260/A280 is in the range of 2.1 or so for control；
2.3 take 600ng purified genomic dna sample, and denaturation treatment is carried out using NaOH solution, obtain being denatured genome
DNA；
2.4 pairs of purified genomic dna samples expand, and obtain genome amplification product；
2.5 use PstI and SacI to obtain sample DNA fragment as the random digestion genome amplification product of the second enzyme liquid；
2.6 pairs of sample DNA fragments carry out single base extensions, carry out Two Colour Fluorescence dye marker, then with red ox DNA
Chip Bovine High Density chip carry out hybridization reaction；
After 2.7 cleaning chips, the fluorescence data of chip is scanned；
2.8 are obtained by analysis, and the related molecular labeling of Red Steppe fat content includes carcass weight molecular labeling, thick fat
Fat molecular labeling and meat weight molecular labeling.
Embodiment 4
The present embodiment, which provides the related molecular labeling of Red Steppe fat content, includes crude fat content molecular labeling, cutability
Weight molecule marks and carcass weight molecular labeling；Crude fat content molecular labeling includes ELAVL2 genetic mutation sites and ELAVL2 bases
Because of variant sites, meat weight molecular labeling includes LOC104970972 genetic mutation sites, RAP2B genetic mutation sites, MBNL1
Genetic mutation site and CCDC30 genetic mutation sites, carcass weight molecular labeling include LTA4H genetic mutation sites,
C7H5orf63 genetic mutation sites, LOC101907891 genetic mutation sites, KCNS3 genetic mutation sites, LOC786289 bases
Because of variant sites and FER1L6 genetic mutation sites.
The LTA4H genetic mutation sites of carcass weight molecular labeling are loc60711494, C7H5orf63 genetic mutation site
It is that loc10665717, KCNS3 genetic mutation site are for loc28343212, LOC101907891 genetic mutation site
Loc80905529, LOC786289 genetic mutation site are that loc60324725 and FER1L6 genetic mutation sites are
Loc17551045, and MED30 genetic mutation sites are loc49023940 and loc49032157.
Experimental example 1
This experimental example provides the acquisition methods of the related molecular labeling of the Red Steppe fat content provided embodiment 1,
And analyze the related molecular labeling of Red Steppe fat content and the correlation of meat index of correlation.
The method provided by embodiment 1 obtains crude fat content molecular labeling, meat weight molecular labeling and carcass weight point
Son mark, specific experiment result is as shown in table 1- tables 3.
The significantly correlated SNP results of the crude fat of the Red Steppe of table 1
The significantly correlated SNP results of the meat weight of the Red Steppe of table 2
The significantly correlated SNP results of the carcass weight of the Red Steppe of table 3
The carcass weight molecular labeling, crude fat molecular labeling and cutability of Red Steppe are can be seen that from the data of table 1- tables 3
Weight molecule marks, and wherein crude fat content molecular labeling includes ELAVL2 genetic mutation sites and ELAVL2 genetic mutation sites,
Meat weight molecular labeling includes LOC104970972 genetic mutation sites, RAP2B genetic mutation sites, MBNL1 genetic mutations position
Point and CCDC30 genetic mutation sites, carcass weight molecular labeling include LTA4H genetic mutation sites, C7H5orf63 genetic mutations
Site, LOC101907891 genetic mutation sites, KCNS3 genetic mutation sites, LOC786289 genetic mutation sites and FER1L6
Genetic mutation site, it is closely related with carcass weight, crude fat and meat weight respectively, in may apply to livestock-raising and butchering.
In summary, the preparation method of the related molecular labeling of Red Steppe fat content provided in an embodiment of the present invention,
Carcass weight molecular labeling, crude fat molecular labeling and meat weight molecular labeling can be included by testing quickly to obtain, wherein slightly
Fat content molecular labeling, there is higher application value.
Embodiments described above is part of the embodiment of the present invention, rather than whole embodiments.The reality of the present invention
The detailed description for applying example is not intended to limit the scope of claimed invention, but is merely representative of the selected implementation of the present invention
Example.Based on the embodiment in the present invention, what those of ordinary skill in the art were obtained under the premise of creative work is not made
Every other embodiment, belongs to the scope of protection of the invention.
Claims (10)
1. the acquisition methods of the related molecular labeling of Red Steppe fat content, it is characterised in that comprise the following steps：
The genomic DNA of Red Steppe is extracted, and with the first enzyme liquid digestion process, obtains purified genomic dna；
The 500-700ng purified genomic dna denaturation treatment is taken, obtains being denatured genomic DNA；
The denaturation genomic DNA is expanded, and obtains genome amplification product；The genome amplification product is through the second enzyme liquid
Processing, obtains sample DNA fragment；
React, and fluorescent scanning and analysis, obtain and Red Steppe with DNA chip after the sample DNA fragment Single base extension
The related molecular labeling of fat content.
2. the acquisition methods of the related molecular labeling of Red Steppe fat content according to claim 1, it is characterised in that institute
The A260/A280 values for stating genomic DNA are 1.8-2.1.
3. the acquisition methods of the related molecular labeling of Red Steppe fat content according to claim 2, it is characterised in that institute
The concentration for stating genomic DNA is 500ng/ μ L-2000ng/ μ L.
4. the acquisition methods of the related molecular labeling of Red Steppe fat content according to claim 1, it is characterised in that institute
It is at least one of RNase or protease to state the first enzyme liquid.
5. the acquisition methods of the related molecular labeling of Red Steppe fat content according to claim 1, it is characterised in that institute
The denaturant for stating denaturation treatment is NaOH solution.
6. the acquisition methods of the related molecular labeling of Red Steppe fat content according to claim 1, it is characterised in that institute
Stating the second enzyme liquid includes restriction endonuclease.
7. the acquisition methods of the related molecular labeling of Red Steppe fat content according to claim 6, it is characterised in that institute
Stating also includes precipitation after restriction endonuclease handles the genome amplification product and is resuspended, the reaction reagent of the precipitation
For ethanol solution.
8. the acquisition methods of the related molecular labeling of Red Steppe fat content according to claim 1, it is characterised in that institute
Stating the related molecular labeling of Red Steppe fat content includes crude fat content molecular labeling, meat weight molecular labeling and carcass weight
Molecular labeling；The crude fat content molecular labeling includes ELAVL2 genetic mutation sites and ELAVL2 genetic mutation sites, institute
Stating meat weight molecular labeling includes LOC104970972 genetic mutation sites, RAP2B genetic mutation sites, MBNL1 genetic mutations
Site and CCDC30 genetic mutation sites, the carcass weight molecular labeling include LTA4H genetic mutation sites, C7H5orf63 bases
Because variant sites, LOC101907891 genetic mutation sites, KCNS3 genetic mutation sites, LOC786289 genetic mutation sites and
FER1L6 genetic mutation sites.
9. the related molecular labeling of Red Steppe fat content, it is characterised in that related point of the Red Steppe fat content
Son mark is obtained by the acquisition methods of the related molecular labeling of the Red Steppe fat content as described in claim any one of 1-8
.
10. the related molecular labeling of Red Steppe fat content as claimed in claim 9 reflects in Red Steppe meat fat content
Application in fixed.
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