CN107828784A - Extracted from complex samples and purify the kit and method of e. coli dna - Google Patents
Extracted from complex samples and purify the kit and method of e. coli dna Download PDFInfo
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- CN107828784A CN107828784A CN201711431675.1A CN201711431675A CN107828784A CN 107828784 A CN107828784 A CN 107828784A CN 201711431675 A CN201711431675 A CN 201711431675A CN 107828784 A CN107828784 A CN 107828784A
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- 108020004707 nucleic acids Proteins 0.000 claims abstract description 30
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 30
- 238000002955 isolation Methods 0.000 claims abstract description 14
- 238000010521 absorption reaction Methods 0.000 claims abstract description 11
- 239000003480 eluent Substances 0.000 claims abstract description 10
- 239000006166 lysate Substances 0.000 claims abstract description 10
- 238000004140 cleaning Methods 0.000 claims abstract description 8
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- 238000000746 purification Methods 0.000 claims abstract description 8
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- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 18
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- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims description 9
- 239000011780 sodium chloride Substances 0.000 claims description 9
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- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 claims description 7
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- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 6
- 238000002156 mixing Methods 0.000 claims description 6
- 101000850928 Mycobacterium phage L5 Gene 37 protein Proteins 0.000 claims description 5
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 5
- 238000007865 diluting Methods 0.000 claims description 4
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 claims description 3
- BTBUEUYNUDRHOZ-UHFFFAOYSA-N Borate Chemical compound [O-]B([O-])[O-] BTBUEUYNUDRHOZ-UHFFFAOYSA-N 0.000 claims description 3
- 241000588722 Escherichia Species 0.000 claims description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 3
- 239000007836 KH2PO4 Substances 0.000 claims description 3
- 238000006243 chemical reaction Methods 0.000 claims description 3
- 238000005336 cracking Methods 0.000 claims description 3
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 claims description 3
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 3
- 229910000397 disodium phosphate Inorganic materials 0.000 claims description 3
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- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims 1
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- SCYULBFZEHDVBN-UHFFFAOYSA-N 1,1-Dichloroethane Chemical class CC(Cl)Cl SCYULBFZEHDVBN-UHFFFAOYSA-N 0.000 description 1
- 241000186146 Brevibacterium Species 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
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- TVWHNULVHGKJHS-UHFFFAOYSA-N Uric acid Natural products N1C(=O)NC(=O)C2NC(=O)NC21 TVWHNULVHGKJHS-UHFFFAOYSA-N 0.000 description 1
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- 229940099352 cholate Drugs 0.000 description 1
- BHQCQFFYRZLCQQ-OELDTZBJSA-N cholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-OELDTZBJSA-N 0.000 description 1
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- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
- C12N15/1013—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers by using magnetic beads
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Abstract
The invention discloses a kind of kit and method for being extracted from complex samples and purifying e. coli dna, belong to biology field.The present invention extracts from complex samples and purifies the kit of e. coli dna, including：1）It has been coupled the carboxylated nanometer magnetic bead Eget103 of gp37 albumen；2）Sample dilution；3）Sample lysate；4）Nucleic acid cleaning fluid；5）DNA eluents 6）Nucleic acid absorption magnetic bead.The present invention is then purified using the carboxylated nanometer magnetic bead Eget103 specific binding Escherichia coli for being coupled gp37 albumen to Escherichia coli, and Escherichia coli after purification then are extracted into DNA with paramagnetic particle method.Isolation and purification culture of the whole process without Escherichia coli, directly carry out DNA extractions；Simplify experimentation, greatly save experimental period.
Description
Technical field
It is more particularly to a kind of to be extracted from complex samples and purify Escherichia coli the present invention relates to biology field
DNA kit and method.
Background technology
EHEC, also known as Escherichia coli（Gram-negative brevibacterium）, it is the common perch in humans and animals enteron aisle
Bacterium.According to pathogenic difference, Diarrhoea-causing Escherichia E.coli is divided into production enterotoxin EHEC, enteron aisle invasion large intestine
Angstrom 5 kinds of uncommon bacterium, enteropathic EHEC, intestines concentration adhesion EHEC and enterohemorrhagic escherichia coli.
T4 bacteriophages are a kind of virus using Escherichia coli as host, and host specificity is very strong.Gp37 albumen is located at T4 classes
The long tailfiber end of bacteriophage, its C-terminal can identify Host Strains surface receptor, trigger phage adsorption to the mistake of surface of E. coli
Journey, it is the key protein of T4 class bacteriophage identification receptors.
Complex samples refer to food, excrement, soil, vomitus equal samples, these samples typically all containing suppress downstream PCR or
The inhibitor of person NGS detections, such as complicated polysaccharide, cholate, lipid and uric acid etc..Tradition is from complex samples（Food, excrement, soil
Earth, vomitus etc.）The method of middle extraction e. coli dna needs first to separate from sample and purify culture bacterial strain, then from purifying
Nucleic acid is extracted in good bacterial strain, process is more complicated, and the time used is longer.
The content of the invention
In order to make up the deficiencies in the prior art, solves the existing procedure that e. coli dna is extracted from complex samples
Complicated, the problem of required time is long, the invention provides a kind of reagent for being extracted from complex samples and purifying e. coli dna
Box and method.
The technical scheme is that：
A kind of kit for being extracted from complex samples and purifying e. coli dna, it is characterised in that including：
1）It has been coupled the carboxylated nanometer magnetic bead Eget103 of gp37 albumen；
2）Sample dilution；
3）Sample lysate；
4）Nucleic acid cleaning fluid；
5）DNA eluents
6）Nucleic acid absorption magnetic bead.
Preferably, step 1）The middle carboxyl nanometer magnetic bead Eget103 for being coupled gp37 albumen preparation method is：
a）Using the escherichia expression system of gp37 albumen, solubility expression gp37 albumen simultaneously purifies；
b）Activated hydroxyl groups nanometer magnetic bead：
c）Gp37 albumen is coupled with hydroxylating nanometer magnetic bead.
Further, step c）The detailed process that gp37 albumen is coupled with hydroxylating nanometer magnetic bead is：
Using the borate solution containing Tween-20 as coupling buffer, hydroxylating nano magnetic is cleaned with the coupling buffer
Pearl；
The hydroxylating nanometer magnetic bead, gp37 albumen after purification and coupling buffer are mixed, 2-6 is coupled at 20-30 DEG C
Hour；Magnetic Isolation removes supernatant, adds PBST solution and magnetic bead, 20-30 DEG C of reaction 30-90min closing magnetic bead surfaces are resuspended
Unreacted activated hydroxyl groups group；
Magnetic Isolation supernatant, after PBS or preservation solution washing, it is placed in and preserves in solution, 4 DEG C of preservations.
Preferably, the pH of the coupling buffer is 8.0-9.0.It is furthermore preferred that the pH of the coupling buffer
For 8.5.
Preferably, the composition of the Sample dilution is：NaCl 137mM, KCl 2.7mM, Na2HPO4
4.3mM, KH2PO41.4mM, 0.05%Tween-20；
The composition of the sample lysate is：NaCl 0.05mM, EDTA 5mM, the Tris-HCl 20mM of PH=8.0, bacteriolyze
Enzyme 3mg/ml.
Preferably, the composition of the nucleic acid cleaning fluid is：NaCl 0.05mM, EDTA 0.05mM, PH=8.0
Tris-HCl 20mM, absolute ethyl alcohol 70%.
Preferably, the composition of the DNA eluents is：EDTA 0.05mM, the Tris-HCl of PH=8.0
10mM。
The method extracted using the kit from complex samples and purify e. coli dna, including step：
1）Complex samples are taken, add the sample diluting liquid, after mixing, centrifuging and taking supernatant；
2）Add the carboxylated nanometer magnetic bead Eget103 for being coupled gp37 albumen, shaken cultivation 10- at 36.5-37.5 DEG C
20min；
3）Magnetic Isolation combines the magnetic bead of Escherichia coli, removes liquid, and the magnetic bead magnetic bead for combining Escherichia coli uses sample
Dilution cleans；
4）Sample lysate is added into the magnetic bead for combine Escherichia coli, 15-20min is incubated at 36.5-37.5 DEG C, is cracked
Thalline；
5）After cracking, room temperature is cooled to, adds nucleic acid absorption magnetic bead, room temperature mixes；
6）The magnetic bead for being adsorbed with nucleic acid is cleaned using nucleic acid cleaning fluid；
7）DNA eluents are added into the magnetic bead for be adsorbed with nucleic acid, are incubated at 36.5-37.5 DEG C, then Magnetic Isolation, supernatant
Preserved at -20 DEG C.
Preferably, step 1）In, 8-10 mL sample diluting liquids are added in 1g complex samples；Step 2）Middle coupling
The carboxylated nanometer magnetic bead Eget103 of gp37 albumen and step 1）The volume ratio of supernatant is 1:0.6-1.5.
Preferably, step 4）In, the volume ratio of the magnetic bead and sample lysate that combine Escherichia coli is 1:3-
5；Step 5）In, the volume ratio of nucleic acid absorption magnetic bead and the magnetic bead for combining Escherichia coli is 2.5-3.5:2；Step 7）In, DNA
The volume ratio of eluent and nucleic acid absorption magnetic bead is 1:5-7.
Beneficial effects of the present invention are：
The present invention specifically binds Escherichia coli then to big using the carboxylated nanometer magnetic bead Eget103 for being coupled gp37 albumen
Enterobacteria is purified, and Escherichia coli after purification then are extracted into DNA with paramagnetic particle method.Point of the whole process without Escherichia coli
Cultivated from purifying, directly carry out DNA extractions；Simplify experimentation, greatly save experimental period.
Brief description of the drawings
In order to illustrate more clearly about the embodiment of the present invention or technical scheme of the prior art, below will be to embodiment or existing
There is the required accompanying drawing used in technology description to be briefly described, it should be apparent that, drawings in the following description are only this
Some embodiments of invention, for those of ordinary skill in the art, without having to pay creative labor, may be used also
To obtain other accompanying drawings according to these accompanying drawings.
Fig. 1 is the electrophoretogram of gp37 albumen after purification；
Fig. 2 is that the soak time of hydroxylating nanometer magnetic bead influences situation map to gp37 albumen couplings amount；
Fig. 3 is that the pH of coupling buffer influences situation map to gp37 albumen couplings amount；
Fig. 4 is influence situation map of the coupling time to gp37 albumen coupling amounts；
Fig. 5 is the design sketch of the inventive method PCR nucleic acid extractions.
Embodiment
Embodiment 1
A kind of kit for being extracted from complex samples and purifying e. coli dna, it is characterised in that including：
1）It has been coupled the carboxylated nanometer magnetic bead Eget103 of gp37 albumen；
2）Sample dilution（Reagent A）；
3）Sample lysate（Reagent B）；
4）Nucleic acid cleaning fluid（Reagent C）；
5）DNA eluents（Reagent D）
6）Nucleic acid absorption magnetic bead；
Wherein,
The composition of reagent A is：NaCl 137mM, KCl 2.7mM, Na2HPO4 4.3mM, KH2PO41.4mM, 0.05%
Tween-20。
Reagent B composition is：NaCl 0.05mM, EDTA 5mM, the Tris-HCl 20mM of PH=8.0, lysozyme
3mg/ml。
The composition of reagent C is：NaCl 0.05mM, EDTA 0.05mM, the Tris-HCl 20mM of PH=8.0, anhydrous second
Alcohol 70%.
The composition of reagent D is：EDTA 0.05mM, the Tris-HCl 10mM of PH=8.0.
The preparation method for being coupled the carboxyl nanometer magnetic bead Eget103 of gp37 albumen is：
a）Using the escherichia expression system of gp37 albumen, solubility expression gp37 albumen simultaneously purifies；Gp37 eggs after purification
White protein electrophoresis figure is as shown in Figure 1.
b）Activated hydroxyl groups nanometer magnetic bead；
Carboxylated nanometer magnetic bead uses the nanometer magnetic bead of commercialization（Suzhou castor biomedical engineering Co., Ltd MagCOOH）,
After mixing magnetic bead, 100 uL Mag COOH magnetic beads are taken into 1.5 mL centrifuge tubes, and Magnetic Isolation removes supernatant, with 200 uL
MEST solution（100 mM MES (2-（N- morpholines）Ethyl sulfonic acid monohydrate）, pH 5.0,0.05% Tween 20）Carry out magnetic
Separating, washing 2 times, then removes supernatant；
It is rapidly added 100 μ L EDC of Fresh（Dichloroethanes）Solution（10 mg/mL, disperseed with above-mentioned MEST solution
Agent）With 100 μ L NHS（10 mg/mL, dispersant is made with above-mentioned MEST solution）Solution is into the centrifuge tube equipped with magnetic bead, whirlpool
Mixing makes magnetic bead fully suspend, and 25 DEG C of 30 min of activation, keeps the suspended state of magnetic bead during this period, is carried out using vertical mixed instrument
It is reverse to mix；After above-mentioned steps, the carboxyl of magnetic bead surfaces is activated, can enter with the bio-ligand with primary amino radical
Row covalent coupling.
The soak time of hydroxylating nanometer magnetic bead is influenceed than more significant on albumen coupling amount, as shown in Fig. 2 when magnetic bead activates
Between most preferably 30min, soak time is long or too short can all influence gp37 albumen coupling amounts.
c）Gp37 albumen is coupled with hydroxylating nanometer magnetic bead.
Step c）The detailed process that gp37 albumen is coupled with hydroxylating nanometer magnetic bead is：
A. Magnetic Isolation removes supernatant, with Tween-20 of the 1mmol/L borate solution containing 0.05g/dL（Abbreviation BST）Make
For coupling buffer, first magnetic bead being cleaned 2 times with BST, adding the gp37 of appropriate BST and 150ug after purification, volume is settled to
300uL；Wherein, the pH of coupling buffer is 8.0-9.0, most preferably 8.5.The pH of coupling buffer is to gp37 albumen coupling amounts
Influence significantly, as shown in figure 3, when coupling buffer pH is 8.5, gp37 albumen couplings amount is maximum.
B.25 DEG C 4 h of coupling, reverse mixing is carried out using vertical mixed instrument；
Coupling time also has considerable influence to gp37 albumen coupling amounts, as shown in figure 4, coupling time is 4 hours, gp37 albumen is even
Connection amount is maximum.
C. centrifuge tube is placed in Magnetic Isolation on magnetic separation rack and removes supernatant, adds 300 uL PBST solution（pH
7.2, and contain 1%BSA）Magnetic bead is resuspended, 25 DEG C of reaction 1 h closing unreacted activated carboxyl groups of magnetic bead surfaces, keeps during this period
The suspended state of magnetic bead.
D. centrifuge tube is placed in Magnetic Isolation on magnetic separator and removes supernatant, every time with 200 uL PBS solutions（pH
7.2）Or after preservation solution washs 3 times, it is resuspended in preserving solution（Containing 0.02g/dLNaN3BST-BSA）In, obtain even
Join the carboxylated nanometer magnetic bead Eget103 of gp37 albumen, be stored in 4 DEG C.
The method extracted using kit from complex samples and purify e. coli dna, including step：
1）1g complex samples are taken, add 9mL reagent As in 50mL centrifuge tube, after fully mixing, centrifugation, take 200 μ L of supernatant liquid
In 1.5mL centrifuge tube；
2）Add the carboxylated nanometer magnetic bead Eget103 that 200 μ L have been coupled gp37 albumen, 37 DEG C, 100rpm shaken cultivations
15min；
3）The magnetic bead that magnetic frame Magnetic Isolation combines Escherichia coli is placed in, removes liquid, combines the magnetic bead magnetic of Escherichia coli
Pearl is cleaned at least 3 times using 400 μ L reagent As；
4）800 μ L reagent B are added into the magnetic bead for combine Escherichia coli, 15-20min is incubated at 37 DEG C, crack thalline；
5）After cracking, centrifuge tube is taken out from incubation equipment, room temperature is cooled to, adds nucleic acid absorption magnetic bead（It is purchased from
BioMag article No.s：BMQ300) 300 μ L, room temperature mix；Centrifuge tube is placed in magnetic separation on magnetic frame, waste liquid is removed and inhales
Net lid and ttom of pipe residual night；
6）The magnetic bead for being adsorbed with nucleic acid is cleaned using 1mL reagent Cs, magnetic separation, the lid that exhausts and ttom of pipe residual night；Repeat above-mentioned step
Suddenly twice, uncap and dry at room temperature；
7）50 μ L reagent Ds are added into the magnetic bead for be adsorbed with nucleic acid, it is anti-terminate in water-bath incubate after magnetic separation, draw supernatant
Liquid preserves in new EP pipes at -20 DEG C.
Fig. 5 is the design sketch that nucleic acid extraction is carried out using the inventive method, and as shown in Figure 5, the present invention can be carried effectively
The nucleic acid of Escherichia coli is taken, and extraction process need not isolate and purify sample, can effectively save from various complex samples
Extract the time of nucleic acid.
Claims (10)
- A kind of 1. kit for being extracted from complex samples and purifying e. coli dna, it is characterised in that including：1）It has been coupled the carboxylated nanometer magnetic bead Eget103 of gp37 albumen；2）Sample dilution；3）Sample lysate；4）Nucleic acid cleaning fluid；5）DNA eluents6）Nucleic acid absorption magnetic bead.
- 2. extracted as claimed in claim 1 from complex samples and purify the kit of e. coli dna, it is characterised in that step Rapid 1）The middle carboxyl nanometer magnetic bead Eget103 for being coupled gp37 albumen preparation method is：a）Using the escherichia expression system of gp37 albumen, solubility expression gp37 albumen simultaneously purifies；b）Activated hydroxyl groups nanometer magnetic bead：c）Gp37 albumen is coupled with hydroxylating nanometer magnetic bead.
- 3. extracted as claimed in claim 2 from complex samples and purify the kit of e. coli dna, it is characterised in that step Rapid c）The detailed process that gp37 albumen is coupled with hydroxylating nanometer magnetic bead is：Using the borate solution containing Tween-20 as coupling buffer, hydroxylating nano magnetic is cleaned with the coupling buffer Pearl；The hydroxylating nanometer magnetic bead, gp37 albumen after purification and coupling buffer are mixed, 2-6 is coupled at 20-30 DEG C Hour；Magnetic Isolation removes supernatant, adds PBST solution and magnetic bead, 20-30 DEG C of reaction 30-90min closing magnetic bead surfaces are resuspended Unreacted activated hydroxyl groups group；Magnetic Isolation supernatant, after PBS or preservation solution washing, it is placed in and preserves in solution, 4 DEG C of preservations.
- 4. extracted as claimed in claim 3 from complex samples and purify the kit of e. coli dna, it is characterised in that institute The pH for stating coupling buffer is 8.0-9.0.
- 5. extracted as claimed in claim 1 from complex samples and purify the kit of e. coli dna, it is characterised in that institute The composition for stating Sample dilution is：NaCl 137mM, KCl 2.7mM, Na2HPO4 4.3mM, KH2PO4 1.4mM, 0.05%Tween-20；The composition of the sample lysate is：NaCl 0.05mM, EDTA 5mM, the Tris-HCl of PH=8.0 20mM, lysozyme 3mg/ml.
- 6. extracted as claimed in claim 1 from complex samples and purify the kit of e. coli dna, it is characterised in that institute The composition for stating nucleic acid cleaning fluid is：NaCl 0.05mM, EDTA 0.05mM, the Tris-HCl 20mM of PH=8.0, anhydrous second Alcohol 70%.
- 7. extracted as claimed in claim 1 from complex samples and purify the kit of e. coli dna, it is characterised in that institute The composition for stating DNA eluents is：EDTA 0.05mM, the Tris-HCl 10mM of PH=8.0.
- 8. the method extracted from complex samples using kit described in claim 1 and purified e. coli dna, its feature exist In, including step：1）Complex samples are taken, add the sample diluting liquid, after mixing, centrifuging and taking supernatant；2）Add the carboxylated nanometer magnetic bead Eget103 for being coupled gp37 albumen, shaken cultivation 10- at 36.5-37.5 DEG C 20min；3）Magnetic Isolation combines the magnetic bead of Escherichia coli, removes liquid, and the magnetic bead magnetic bead for combining Escherichia coli uses sample Dilution cleans；4）Sample lysate is added into the magnetic bead for combine Escherichia coli, 15-20min is incubated at 36.5-37.5 DEG C, is cracked Thalline；5）After cracking, room temperature is cooled to, adds nucleic acid absorption magnetic bead, room temperature mixes；6）The magnetic bead for being adsorbed with nucleic acid is cleaned using nucleic acid cleaning fluid；7）DNA eluents are added into the magnetic bead for be adsorbed with nucleic acid, are incubated at 36.5-37.5 DEG C, then Magnetic Isolation, supernatant Preserved at -20 DEG C.
- 9. the method extracted as claimed in claim 8 from complex samples and purify e. coli dna, it is characterised in that：Step 1）In, 8-10 mL sample diluting liquids are added in 1g complex samples；Step 2）The middle carboxylated nanometer magnetic bead for being coupled gp37 albumen Eget103 and step 1）The volume ratio of supernatant is 1:0.6-1.5.
- 10. the method extracted as claimed in claim 8 from complex samples and purify e. coli dna, it is characterised in that：Step 4）In, the volume ratio of the magnetic bead and sample lysate that combine Escherichia coli is 1:3-5；Step 5）In, nucleic acid absorption magnetic bead with The volume ratio for combining the magnetic bead of Escherichia coli is 2.5-3.5:2；Step 7）In, the body of DNA eluents and nucleic acid absorption magnetic bead Product is than being 1:5-7.
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| CN109265566A (en) * | 2018-10-16 | 2019-01-25 | 生工生物工程（上海）股份有限公司 | Purification process, kit and its application of 6 × HIS fusion protein |
| CN111218527A (en) * | 2020-03-10 | 2020-06-02 | 广州赛百纯生物科技有限公司 | Environment sample African swine fever virus detection kit and detection method |
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Denomination of invention: Kit and method for extracting and purifying Escherichia coli DNA from complex samples Granted publication date: 20210323 Pledgee: Jinan Chengxi sub branch of Qilu Bank Co.,Ltd. Pledgor: SHANDONG KAIJING BIOTECHNOLOGY CO.,LTD. Registration number: Y2024980012924 |