CN110438119A - A kind of extracting method of crude oil microbe genome DNA - Google Patents
A kind of extracting method of crude oil microbe genome DNA Download PDFInfo
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- CN110438119A CN110438119A CN201910782148.8A CN201910782148A CN110438119A CN 110438119 A CN110438119 A CN 110438119A CN 201910782148 A CN201910782148 A CN 201910782148A CN 110438119 A CN110438119 A CN 110438119A
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- crude oil
- oil phase
- genome dna
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
Abstract
The invention discloses a kind of extracting methods of crude oil oil phase microbe genome DNA.The extracting method of crude oil oil phase microbe genome DNA provided by the invention, comprising: 1) organic compound is mixed with crude oil oil phase sample, obtain mixture, organic compound is isooctane, normal heptane, toluene or carbon disulfide；2) microorganism in separating mixture obtains crude oil oil phase microorganism；3) crude oil oil phase microbe genome DNA is extracted, microbe genome DNA in crude oil oil phase sample is obtained.The requirement of sequencing is able to satisfy using the obtained crude oil oil phase microbe genome DNA of extracting method of crude oil oil phase microbe genome DNA of the invention, DNA concentration > 10ng/ μ l, OD260/280 is between 1.80-2.00, and gained genomic DNA all has good integrality, can be used for extracting crude oil oil phase microbe genome DNA.
Description
Technical field
The present invention relates in field of biotechnology, a kind of extracting method of crude oil microbe genome DNA.
Background technique
Microbe oil production is since its is at low cost, effect is good, have wide range of applications, environmentally protective feature is by Global Oil
The extensive concern of exploration, exploitation.Microbial Enhanced Oil Recovery is mainly that the degradation of microorganism itself and metabolite is utilized to make
Improve oil property for crude oil to improve oil recovery factor.Oily phase microorganism in crude oil is the core of Microbial Enhanced Oil Recovery
The heart, thus it is very important for the understanding of the biological community structure of crude oil.Due to the limitation of current microbe separation technology,
Only 1% microorganism can be isolated and cultured at present, and scientific research personnel develops the second generation high throughput for exempting from culture thus
Sequencing technologies are sequenced by the genomic fragment in extraction environment sample using amplification rRNA hypervariable region, thus
The objective Bacterial community and abundance ratio for obtaining sample itself.And current method is only with respect to Produced Liquid and the base of grease phase
Because a group DNA is extracted, there is no the individually extracting methods about crude oil oil phase microorganism, and crude oil itself is a petroleum
High hydrophobicity, the microorganism of oily Xiangli can survive because of its unique living environment in this complex environment of oil reservoir
And play degradation.And the crude oil oil phase microbe genome DNA for meeting sequencing and requiring can be obtained currently without method, institute
To be badly in need of a kind of method that can extract crude oil oil phase microorganism, and the genomic DNA obtained meets wanting for high-flux sequence
It asks.
Summary of the invention
The object of the present invention is to provide a kind of extracting methods of crude oil oil phase microbe genome DNA.
The extracting method of crude oil oil phase microbe genome DNA provided by the invention, including 1), 2) He 3):
1) organic compound is mixed with crude oil oil phase sample, obtains mixture；The organic compound is isooctane, just
Heptane, toluene or carbon disulfide；
2) microorganism in the mixture is separated, crude oil oil phase microorganism is obtained；
3) the crude oil oil phase microbe genome DNA is extracted, microbial genome in the crude oil oil phase sample is obtained
DNA。
The organic compound can be sterile organic compound.The sterile organic compound can be by by organic compound
0.22 μm of sterile organic phase filter is crossed to obtain.
In the above method, the volume ratio of the organic compound and the crude oil oil phase sample can be 2:1-10:1.It is described
Numerical value of the viscosity of crude oil oil phase under 30 degree is 50-220000mPas).
Further, the volume ratio of the organic compound and the crude oil oil phase sample can be 4:1-8:1.
In the above method, the volume ratio of the organic compound and the crude oil oil phase sample can be 6:1.
In the above method, step 2) can include: the mixture is crossed into organic phase miillpore filter, it is micro- to collect the organic phase
Filter residue on the filter membrane of hole obtains the crude oil oil phase microorganism.
The organic phase miillpore filter can be sterilised membrane filter.
In the above method, the mixture, which is crossed organic phase miillpore filter, to be completed using circulating vacuum water pump.
In the above method, organic phase miillpore filter aperture can be 0.22 μm.
In the above method, edaphon genome is can be used in the extraction crude oil oil phase microbe genome DNA
DNA extraction method is completed.
In the above method, edaphon genome is can be used in the extraction crude oil oil phase microbe genome DNA
DNA extraction kit is completed.
In one embodiment of the invention, the soil microbe genome DNA extracts kit is U.S. MP
The kit that the article No. of Biomedicals company is 116560200.
In one embodiment of the invention, step 2) includes: and receives the excessively described organic phase miillpore filter of the mixture
Collect the organic phase miillpore filter containing filter residue, obtains the filter membrane for being loaded with the crude oil oil phase microorganism；
Step 3) includes: to shred the filter membrane for being loaded with the crude oil oil phase microorganism, obtains filter membrane fragment；Using described
Soil microbe genome DNA extracts kit extracts the genomic DNA of microorganism on the filter membrane fragment, obtains the crude oil
Microbe genome DNA in oily phase sample.
Sample treatment before being extracted the present invention also provides crude oil oil phase microbe genome DNA, which comprises
Organic compound is mixed to the processing for realizing sample with crude oil oil phase sample；The organic compound is isooctane, normal heptane, first
Benzene or carbon disulfide.
The present invention also provides one group of complete sets of products, the complete sets of products can be following X1), X2) or X3):
X1) reagent by organic compound and for extracting microbe genome DNA or the complete sets of products of kit forms；
The organic compound is isooctane, normal heptane, toluene or carbon disulfide；
X2) by the organic compound, the reagent for extracting microbe genome DNA or kit and organic phase micropore
The complete sets of products of filter membrane composition；
X3) by the organic compound, the reagent for extracting microbe genome DNA or kit, organic phase micropore
The complete sets of products of filter membrane and circulating vacuum water pump composition.
The production set product can be used for extracting crude oil oil phase microbe genome DNA.
The reagent or kit for extracting microbe genome DNA can be microorganism base in the non-crude oil oil phase of extraction
Because of reagent used in group DNA or kit.The reagent or kit for extracting microbe genome DNA is concretely
The text soil microbe genome DNA extracts kit.
The extracting method of crude oil oil phase microbe genome DNA microbial community structure in detection crude oil oil phase
And/or the application in abundance, also belong to protection scope of the present invention.
Application of the extracting method of the crude oil oil phase microbe genome DNA in identifying crude oil oil phase in microorganism,
Also belong to protection scope of the present invention.
Application of the complete sets of products in detection crude oil oil phase in microbial community structure and/or abundance, also belongs to this
The protection scope of invention.
Application of the complete sets of products in identifying crude oil oil phase in microorganism, also belongs to protection scope of the present invention.
In the present invention, the organic phase filter is the filter for separating microorganism and organic phase in organic matter.
The organic phase miillpore filter is the filter membrane for separating microorganism and organic phase in organic matter.
Utilize the obtained crude oil oil phase microorganism of the extracting method of crude oil oil phase microbe genome DNA provided by the invention
Genomic DNA is able to satisfy the requirement of high-flux sequence, DNA concentration > 10ng/ μ l, OD260/280 between 1.80-2.00, and
Gained genomic DNA all has good integrality.Show mentioning for crude oil oil phase microbe genome DNA provided by the invention
It takes method to can be used for extracting crude oil oil phase microbe genome DNA, further can be also used for analysis crude oil oil phase microorganism
The identification of group structure and abundance and microorganism, is with a wide range of applications.
Detailed description of the invention
Fig. 1 is the electrophoresis detection result of microbe genome DNA obtained by six parts of crude oil samples.Swimming lane 1Kb indicates DNA molecular
Amount standard, from top to bottom the size of band is followed successively by 10000,8000,6000,5000,4000,3000,2000 and 1000bp, swimming
Road 1-6 is respectively microbe genome DNA obtained by XJ-1, XJ-2, XJ-3, JD-1, JD-2 and JD-3.
Fig. 2 is the electrophoresis detection result that crude oil sample microbe genome DNA is extracted using different organic compounds.Swimming lane
1Kb indicates DNA molecular amount standard, from top to bottom the size of band be followed successively by 10000,8000,6000,5000,4000,3000,
2000 and 1000bp, swimming lane 5-9 are respectively isooctane, normal heptane, control group, toluene, carbon disulfide.
Fig. 3 is the electrophoresis detection result that crude oil sample microbe genome DNA is extracted using different amounts of isooctane.Swimming lane
1Kb indicates DNA molecular amount standard, from top to bottom the size of band be followed successively by 10000,8000,6000,5000,4000,3000,
The volume that 2000 and 1000bp, swimming lane Z1-Z5 respectively indicate isooctane is 20ml, 40ml, 60ml, 80ml, 100ml.
Specific embodiment
The present invention is further described in detail With reference to embodiment, and the embodiment provided is only for explaining
The bright present invention, the range being not intended to be limiting of the invention.Experimental method in following embodiments is unless otherwise specified
Conventional method.Material as used in the following examples, reagent, instrument etc., are commercially available unless otherwise specified.
Quantitative test in following embodiment, is respectively provided with three repeated experiments, and results are averaged.
The extraction of microbe genome DNA in embodiment 1, crude oil
The present embodiment is extracted microbe genome DNA in crude oil, six parts of crude oil samples of microbe genome DNA to be extracted
Product are respectively derived from 3 mouthfuls of wells of 3 mouthfuls of wells (sample number into spectrum XJ-1, XJ-2 and XJ-3) and NW Hebei in Xinjiang, and (sample number into spectrum is
JD-1, JD-2 and JD-3), each well portion sample.
1, microbe genome DNA in crude oil is extracted using isooctane
The extracting method of microbe genome DNA includes the following steps: in every part of sample
The crude oil sample for taking 10ml, the sterile isooctane of 60ml is added, and (sterile isooctane is sterile organic with 0.22 μm
Obtained after the filtering of phase filter), mixture is obtained after mixing；Gained mixture is passed through into circulation vacuum pump (upper sea cowry logical sequence instrument
Device equipment Co., Ltd SHZ-D (III) vacuum pump using circulatory water) 0.22 μm of sterile organic phase miillpore filter is crossed, filtrate is abandoned, collects and cuts
There are organic filter membranes of microorganism；There is organic filter membrane of microorganism to be folded with aseptic nipper retention, with sterile scissors (75% second
Alcohol solution disinfection) shred after be put into 2ml sterile centrifugation tube, obtain pretreatment sample；Then it utilizesSpin
Kit for Soil (soil genome DNA extracting reagent kit, MP Biomedicals company, the U.S., article No. 116560200)
Pretreatment sample genomic DNA is extracted, extraction step is carried out according to kit specification, and obtains the microorganism of six parts of crude oil samples
Genomic DNA.
The concentration and purity (table 1) of gained genomic DNA are detected using NanoDrop2000, and utilize Ago-Gel electricity
The integrality (Fig. 1) of swimming detection gained genomic DNA.
The testing result of table 1,6 part of crude oil sample microbe genome DNA concentration and purity
| Sample number into spectrum | Concentration (ng/ μ l) | OD260/280 | OD260/230 |
| XJ-1 | 85.80 | 1.89 | 0.04 |
| XJ-2 | 190.40 | 1.91 | 0.04 |
| XJ-3 | 77.30 | 1.86 | 0.10 |
| JD-1 | 60.00 | 1.87 | 0.06 |
| JD-2 | 50.80 | 1.92 | 0.11 |
| JD-3 | 100.20 | 1.88 | 0.04 |
Requirement of the current two generations high-flux sequence for the concentration of DNA: DNA concentration > 10ng/ μ l, OD260/280 exist
Between 1.80-2.00, the results show that the concentration and purity of the genomic DNA obtained by the above method are all satisfied two generation high passes
The requirement of sequence is measured, and electrophoresis result is also shown, gained genomic DNA all has good integrality.
2, microbe genome DNA in crude oil is extracted using other organic compounds
According to the method for step 1, isooctane is replaced with to organic compound-normal heptane, toluene, carbon disulfide respectively,
His step is constant, extracts the genome for the crude oil sample that 10ml number is XJ-1 respectively using normal heptane, toluene, carbon disulfide
Then DNA is used the concentration and purity (table 2) for being detected gained genomic DNA using NanoDrop2000, and utilizes Ago-Gel
The integrality (Fig. 2) of genomic DNA obtained by electrophoresis detection.
Control group be with reference to CN102732504A it is a kind of from oil/gas hide environment in extract the macro genome of microorganism method,
Petroleum ether/n-hexane of ethanol water and 5ml volume that 30% (V/V) of 10ml volume is added in 10ml crude oil sample is mixed
Object (volume ratio of petroleum ether and n-hexane is 1:1 in petroleum ether/hexane mixture) is closed, is sufficiently shaken up, it is static to make grease phase
Layering extracts lower layer's water phase with sebific duct, then water phase Crude Oil and other impurities is filtered out with filter paper, finally with 0.22 μm of water system
Filter membrane vacuum filtration is enriched with thallus on filter membrane, filter membrane is folded with aseptic nipper, with sterile scissors (75% ethanol water
Disinfection) shred after be put into 2ml sterile centrifugation tube, obtain pretreatment sample；Then it utilizesSpin Kit for
Soil (soil genome DNA extracting reagent kit, MP Biomedicals company, the U.S., article No. 116560200) extracts pre- place
Sample gene group DNA is managed, extraction step is carried out according to kit specification, and obtains control group microbe genome DNA.Then it uses
The concentration and purity (table 2) of gained genomic DNA are detected using NanoDrop2000, and detect institute using agarose gel electrophoresis
Obtain the integrality (Fig. 2) of genomic DNA.
The results show that the extraction effect of normal heptane, toluene, carbon disulfide is not so good as isooctane, but the genomic DNA obtained
It is also to meet the requirement of two generation high-flux sequences, and the method that control group provides obtains genomic DNA concentration and purity is unsatisfactory for
The requirement of high-flux sequence and electrophoresis result also can't see bright band.Therefore the method for control group can not obtain in crude oil
Oily phase microbe genome DNA.
Table 2, different organic compounds extract the testing result of microbe genome DNA concentration and purity
| Solvent | Concentration (ng/ μ l) | OD260/280 | OD260/230 |
| Isooctane | 87.26 | 1.91 | 0.11 |
| Normal heptane | 14.36 | 1.89 | 0.05 |
| Control group | 3.56 | 1.75 | 0.16 |
| Toluene | 75.76 | 1.87 | 0.08 |
| Carbon disulfide | 84.32 | 1.87 | 0.10 |
3, microbe genome DNA in crude oil is extracted using other organic compounds
According to the method for step 1, by the volume of isooctane by " 60ml " replace with respectively " 20ml ", " 40ml ", " 60ml ",
" 80ml " and " 100ml ", other steps are constant, extract the crude oil that 10ml number is XJ-1 respectively using different amounts of isooctane
The genomic DNA of sample, the then concentration and purity (table 3) of the genomic DNA obtained by being detected using NanoDrop2000, and benefit
The integrality (Fig. 3) of gained genomic DNA is detected with agarose gel electrophoresis.The results show that the genomic DNA obtained is full
The requirement of two generation high-flux sequences of foot.
The testing result of table 3, different sample microbial genomic DNA concentration and purity
| Number | Ratio | Concentration (ng/ μ l) | OD260/280 | OD260/230 |
| Z1 | 2:1 | 15.20 | 1.89 | 0.12 |
| Z2 | 4:1 | 24.30 | 1.90 | 0.06 |
| Z3 | 6:1 | 86.52 | 1.87 | 0.08 |
| Z4 | 8:1 | 104.23 | 1.91 | 0.11 |
| Z5 | 10:1 | 118.62 | 1.88 | 0.07 |
In table 3, data indicate the volume ratio of isooctane and crude oil sample in " ratio " column.
Claims (10)
1. the extracting method of crude oil oil phase microbe genome DNA, including 1), 2) He 3):
1) organic compound is mixed with crude oil oil phase sample, obtains mixture；The organic compound is isooctane, positive heptan
Alkane, toluene or carbon disulfide；
2) microorganism in the mixture is separated, crude oil oil phase microorganism is obtained；
3) the crude oil oil phase microbe genome DNA is extracted, microbe genome DNA in the crude oil oil phase sample is obtained.
2. according to the method described in claim 1, it is characterized by: the body of the organic compound and the crude oil oil phase sample
Product is than being 2:1-10:1；
Further, the volume ratio of the organic compound and the crude oil oil phase sample is 4:1-8:1.
3. method according to claim 1 or 2, it is characterised in that: the organic compound and the crude oil oil phase sample
Volume ratio be 6:1.
4. method according to claim 1 to 3, it is characterised in that: step 2) includes: to have crossed the mixture
Machine phase miillpore filter collects filter residue on the organic phase miillpore filter, obtains the crude oil oil phase microorganism.
5. according to the method described in claim 4, it is characterized by: the mixture is crossed organic phase miillpore filter using circulation
Vacuum pump is completed.
6. method according to claim 4 or 5, it is characterised in that: organic phase miillpore filter aperture is 0.22 μm.
7. any method in -6 according to claim 1, it is characterised in that: described to extract the crude oil oil phase microorganism base
Because group DNA is completed using soil microbe genome DNA extracting method.
Sample treatment before 8. crude oil oil phase microbe genome DNA extracts, comprising: by organic compound and crude oil oil phase sample
The processing of sample is realized in product mixing；The organic compound is isooctane, normal heptane, toluene or carbon disulfide.
9. complete sets of products, for following X1), X2) or X3):
X1) reagent by organic compound and for extracting microbe genome DNA or the complete sets of products of kit forms；It is described
Organic compound is isooctane, normal heptane, toluene or carbon disulfide；
X2) by the organic compound, the reagent for extracting microbe genome DNA or kit and organic phase miillpore filter
The complete sets of products of composition；
X3) by the organic compound, the reagent for extracting microbe genome DNA or kit, organic phase miillpore filter
With the complete sets of products of circulating vacuum water pump composition.
10. any the method answering in microbial community structure and/or abundance in detection crude oil oil phase in claim 1-8
With；
Or, application of any the method in identifying crude oil oil phase in microorganism in claim 1-8；
Or, application of the complete sets of products described in claim 9 in detection crude oil oil phase in microbial community structure and/or abundance；
Or, application of the complete sets of products described in claim 9 in identifying crude oil oil phase in microorganism.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102732504A (en) * | 2011-04-15 | 2012-10-17 | 大连百奥泰科技有限公司 | Method for extracting microorganism macrogenome from oil/gas pool environment |
| CN103667255A (en) * | 2013-11-19 | 2014-03-26 | 克拉玛依市金山石油化工有限公司 | DNA extracting method for petroleum microorganism in raw petroleum environmental sample |
| CN104630204A (en) * | 2013-11-15 | 2015-05-20 | 中国石油化工股份有限公司 | Extraction method of oil reservoir microbial genome DNA |
| CN106754895A (en) * | 2017-03-01 | 2017-05-31 | 中国石油大学(北京) | Extract the method and kit of crude oil total dna |
-
2019
- 2019-08-23 CN CN201910782148.8A patent/CN110438119A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102732504A (en) * | 2011-04-15 | 2012-10-17 | 大连百奥泰科技有限公司 | Method for extracting microorganism macrogenome from oil/gas pool environment |
| CN104630204A (en) * | 2013-11-15 | 2015-05-20 | 中国石油化工股份有限公司 | Extraction method of oil reservoir microbial genome DNA |
| CN103667255A (en) * | 2013-11-19 | 2014-03-26 | 克拉玛依市金山石油化工有限公司 | DNA extracting method for petroleum microorganism in raw petroleum environmental sample |
| CN106754895A (en) * | 2017-03-01 | 2017-05-31 | 中国石油大学(北京) | Extract the method and kit of crude oil total dna |
Non-Patent Citations (2)
| Title |
|---|
| ATHENIA L OLDHAM等: "Automated DNA extraction platforms offer solutions to challenges of assessing microbial biofouling in oil production facilities", 《AMB EXPRESS》 * |
| 任国领等: "大庆油田油藏微生物基因组DNA的提取与分析", 《大庆石油学院学报》 * |
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