CN106198479A - The SYBR Green I immue quantitative detection reagent box of a kind of detection by quantitative DNA content and detection method thereof - Google Patents
The SYBR Green I immue quantitative detection reagent box of a kind of detection by quantitative DNA content and detection method thereof Download PDFInfo
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- CN106198479A CN106198479A CN201610737798.7A CN201610737798A CN106198479A CN 106198479 A CN106198479 A CN 106198479A CN 201610737798 A CN201610737798 A CN 201610737798A CN 106198479 A CN106198479 A CN 106198479A
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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Abstract
The present invention relates to SYBR Green I immue quantitative detection reagent box and the detection method thereof of a kind of detection by quantitative DNA content, including box body, support, an EP pipe and the 2nd EP pipe, wherein, in described box body, support is set；Described support is provided with 46 pipe supports；A described EP pipe is arranged on the first pipe support, is provided with boring, is provided with qualitative filter paper in pipe bottom a described EP pipe；Described 2nd EP pipe is arranged on the second pipe support, is provided with boring, is provided with qualitative filter paper in pipe bottom described 2nd EP pipe.The inventive method is easy and simple to handle, with low cost, accuracy is high.
Description
Technical field
The present invention relates to biochemistry and biology field, be specifically related to the SYBR of a kind of detection by quantitative DNA content
Green I immue quantitative detection reagent box and detection method thereof, it is adaptable to the detection by quantitative of various DNA.
Background technology
Usually need when biology field carries out DNA analysis DNA is carried out accurate quantitative analysis.In clinical diagnosis side
Face, the detection by quantitative of viral DNA is the important evidence of clinical diagnosis.
SYBR Green I is a kind of dye with green excitation wavelength being incorporated into all dsDNA minor groove regions
Material.Under free state, SYBR Green I sends faint fluorescence, but after being once combined with double-stranded DNA, fluorescence increases greatly
By force.Therefore, the fluorescence signal intensity of SYBR Green I is relevant to the quantity of double-stranded DNA, can detect according to fluorescence signal
The double-stranded DNA quantity that PCR system exists.The maximum absorption wavelength of SYBR Green I is about 497nm, launches wavelength and is about
520nm。
SYBR Green I nucleic acid gel dye liquor is the fluorescence dye liquor of a kind of hypersensitivity, can be used for detecting agarose gel
And the double-stranded DNA in polyacrylamide gel and oligonucleotide.After dye liquor is combined with nucleic acid, SYBR Green I nucleic acid gel
The fluorescence signal of dye liquor can be remarkably reinforced, and it is the best that the gel therefore dyeed through SYBR Green I nucleic acid gel dye liquor demonstrates
Cost performance, there is no background fluorescence.
At present, routine utilizes the method for SYBR Green I detection of fluorescent dyes DNA content to there is the deficiency of following aspect:
1, there is non-specific amplification and lead in the method that in PCR amplification procedure, SYBR Green I fluorescent dye is combined with DNA
Cause quantitative inaccurate risk；
2, SYBR Green I fluorescent dye directly acts on sample existence to sample content, detecting instrument sensitivity requirement
High problem, testing cost is higher, and in sample, testing result also can be caused certain impact by other impurity, causes the most not
Accurately.
Summary of the invention
For above prior art problem, it is an object of the invention to provide the SYBR of a kind of detection by quantitative DNA content
Green I immue quantitative detection reagent box and detection method thereof, discharged in sample other impurity and non-specific amplification product to
The impact of whole testing result, more accurately, the detection by quantitative completing DNA content of low cost, comprises the following steps: coarse filtration；Agar
Sugar gel electrophoresis；DNA reclaims；SYBR Green I fluorescence staining；Detection and interpretation of result.The method utilizes the DNA simplified to return
Receiving method obtains highly purified DNA, then by SYBR Green I dyeing and UV spectrophotometer measuring fluorescence, computational analysis
Obtain DNA concentration.Concrete technical scheme is as follows:
The SYBR Green I immue quantitative detection reagent box of a kind of detection by quantitative DNA content, including box body, support, an EP
Pipe and the 2nd EP pipe, wherein, arrange support in described box body；Described support is provided with 4-6 pipe support；A described EP pipe sets
It is placed on the first pipe support, is provided with boring bottom a described EP pipe, in pipe, is provided with qualitative filter paper；Described 2nd EP pipe is arranged at
On two pipe supports, it is provided with boring bottom described 2nd EP pipe, in pipe, is provided with qualitative filter paper.
Further, also include that lid, described lid side are connected to box body and for fastening box body.
Further, a described EP pipe is the EP pipe of 0.5ml or 1.5ml, and described 2nd EP pipe is 0.5ml or 1.5ml
EP pipe.
The detection method of the SYBR Green I immue quantitative detection reagent box of above-mentioned detection by quantitative DNA content, including walking as follows
Rapid:
(1) coarse filtration；
(2) agarose gel electrophoresis；
(3) DNA reclaims；
(4) SYBR Green I fluorescence staining；
(5) detection；
(6) interpretation of result.
Further, step (1) including: (1-1) take testing sample put into be drilled hole in bottom and put into fritter fixed
In the EP pipe of the 0.5ml of property filter paper, then the EP pipe of this 0.5ml is inserted in the EP pipe of a 1.5ml；(1-2) will package
EP pipe is centrifugal, removes the EP pipe of 0.5ml, and the EP of 1.5ml is in control liquid all carries out agarose gel electrophoresis.
Further, step (2) including: (2-1) joins glue；(2-2) electrophoresis prepares；(2-3) loading；(2-4) electrophoresis.
Further, step (3) including: the chopping of the blob of viscose containing target DNA that (3-1) will scale off, and puts into the end of at
Portion is drilled in the EP pipe of hole the 0.5ml that puts into fritter qualitative filter paper, then the EP pipe of this 0.5ml is inserted in a 1.5ml's
In EP pipe；(3-2) the EP pipe packaged is centrifuged, removes the EP pipe of 0.5ml, and the EP of 1.5ml is in control liquid capacity-fixed；(3-
3) NaAc and dehydrated alcohol are added, freezing；Precipitation is dissolved in TE solution after drying.
Further, step (4) including: (4-1) adds the SYBR Green good with the DNA redissolution isopyknic dilution of liquid
I working solution, mixing；(4-2) about 30 minutes are placed.
Further, step (5) including: the DNA solution dyeed is placed on ultraviolet spectrophotometer by (5-1),
Fluorescent value A260 is read respectively at 260nm wavelength；(5-2) according to WdsDNAIn the μ g/ml formula scales testing sample of=A260 × 50
Target DNA content.
Further, (2-1) joins glue: according to genomic fragment size, the agarose gel of configuration respective concentration；Take appropriate
Agar and TAE buffer in conical flask, shake up and seal with tinfoil, put into microwave oven and boil, well-done gel answer bubble-free,
Uniform color；Well-done gel is put in the water-bath of 65 DEG C, when gel is cooled to 65 DEG C, add appropriate SYBR
Green I fluorescent dye, pours into gel in the clean electrophoresis mould installed in time；After gel solidifies completely, careful general
Comb is extracted；(2-2) electrophoresis prepare: gel is put into electrophoresis tank centre together with glue torr, the direction in glue hole towards negative pole,
A certain amount of TAE buffer is poured into just being flooded by gel in electrophoresis tank；(2-3) loading: sample, 6 × bromophenol blue are pressed
The ratio mixing of 5:1, according to the order loading of regulation after gently getting rid of；(2-4) electrophoresis: after loading, covers electrophoresis tank lid, connects electrophoresis
Instrument and electrophoresis tank, arrange electrophoretic parameters: 120V 30min.
The inventive method is easy and simple to handle, with low cost, accuracy is high.
Accompanying drawing explanation
Fig. 1 is the electrophoretogram obtained after electrophoresis of the present invention
Fig. 2 is to be drilled hole in bottom and put into the EP pipe schematic diagram of 0.5ml of fritter qualitative filter paper
Fig. 3 be inserted in be drilled in bottom hole and put into fritter qualitative filter paper 0.5ml EP 1.5ml EP pipe signal
Figure
Fig. 4 is the SYBR Green I immue quantitative detection reagent box of a kind of detection by quantitative DNA content
In figure: swimming lane 1,2 is testing sample A, swimming lane 3,4 is DNA standard control
Detailed description of the invention
Describing the present invention below according to accompanying drawing, it is that the one in numerous embodiments of the present invention is the most real
Execute example.
In a preferred embodiment, a kind of detection by quantitative DNA content SYBR Green I immue quantitative detection reagent box and
Its detection method, comprises the following steps further:
(1) coarse filtration:
1. taking testing sample 100 μ l, the EP putting into the 0.5ml being drilled hole in bottom and putting into fritter qualitative filter paper manages
In, then the EP pipe of this 0.5ml is inserted in the EP pipe of a 1.5ml；
2. the EP pipe centrifugal 10min under 4 DEG C of 10000rpm that will package, removes the EP pipe of 0.5ml, and by the EP of 1.5ml
It is in control liquid and all carries out agarose gel electrophoresis；
(2) agarose gel electrophoresis:
1. glue is joined: according to genomic fragment size, the agarose gel of configuration respective concentration.Take appropriate agar and TAE
Buffer, in conical flask, shakes up and seals with tinfoil, put into microwave oven and boil, and well-done gel answers bubble-free, uniform color.Will
Well-done gel is put in the water-bath of 65 DEG C, when gel is cooled to 65 DEG C, adds appropriate SYBR Green I fluorescence dye
Material, pours into gel in the clean electrophoresis mould installed in time.After gel solidifies completely, careful extracts comb；
2. electrophoresis prepares: gel is put into electrophoresis tank centre together with glue torr, and the direction in glue hole is towards negative pole, toward electricity
Swimming groove pours a certain amount of TAE buffer into just being flooded by gel；
3. loading: sample, 6 × bromophenol blue are mixed in the ratio of 5:1, according to the order loading of regulation after gently getting rid of；
4. electrophoresis: after loading, covers electrophoresis tank lid, connects electrophresis apparatus and electrophoresis tank, arranges electrophoretic parameters: 120V
30min；
(3) DNA reclaims:
1. the chopping of the blob of viscose containing target DNA that will scale off, puts into and is drilled hole in bottom and puts into fritter qualitative filter paper
0.5ml EP pipe in, then the EP pipe of this 0.5ml is inserted in the EP pipe of a 1.5ml；
2. the EP pipe centrifugal 10min under 4 DEG C of 10000rpm that will package, removes the EP pipe of 0.5ml, and by the EP of 1.5ml
It is in control liquid liquid-transfering gun constant volume；
3. 3mol/L NaAc (pH 5.5) and the dehydrated alcohol of 2 times of volumes of 1/10 volume are added ,-20 DEG C of freezings
More than 30min.Precipitation is dissolved in 20 μ l TE solution (pH 7.8) after drying.
(4) SYBR Green I fluorescence staining:
1. (dilution ratio is according to SYBR to add the SYBR Green I working solution good with the DNA redissolution isopyknic dilution of liquid
The description of Green I), mixing；
2. place 30 minutes.
(5) detection and interpretation of result:
1. the DNA solution dyeed is placed on ultraviolet spectrophotometer, at 260nm wavelength, reads fluorescent value respectively
A260；
2. according to WdsDNATarget DNA content in the μ g/ml formula scales testing sample of=A260 × 50.
1. coarse filtration step takes testing sample 100 μ l, puts into and is drilled hole in bottom and puts into the 0.5ml of fritter qualitative filter paper
EP pipe in, then the EP pipe of this 0.5ml is inserted in the EP pipe of a 1.5ml；DNA recycling step 1. by scale off containing mesh
The blob of viscose chopping of DNA, put in the EP pipe that bottom is drilled hole the 0.5ml that puts into fritter qualitative filter paper, then by this
The EP pipe of 0.5ml is inserted in the EP pipe of a 1.5ml；2. the EP pipe packaged is centrifuged under 4 DEG C of 10000rpm by DNA recycling step
5～10min, remove the EP pipe of 0.5ml, and the EP of 1.5ml is in control liquid liquid-transfering gun constant volume；3. DNA recycling step adds
Enter 3mol/L NaAc (pH 5.5) and the dehydrated alcohol of 2 times of volumes of 1/10 volume, at-20 DEG C of freezing more than 30min.Precipitation
It is dissolved in 20 μ l TE solution (pH 7.8) after drying.
In a further advantageous embodiment, a kind of method of SYBR Green I fluorescent dye detection by quantitative DNA content, logical
Cross coarse filtration, agarose gel electrophoresis, DNA recovery, SYBR Green I fluorescence staining, detection and interpretation of result and complete testing sample
The detection by quantitative of DNA, specifically includes following steps:
1, coarse filtration:
(1) taking testing sample 100 μ l, the EP putting into the 0.5ml being drilled hole in bottom and putting into fritter qualitative filter paper manages
In, then the EP pipe of this 0.5ml is inserted in the EP pipe of a 1.5ml；
(2) the EP pipe packaged is centrifuged 5～10min under 4 DEG C of 10000rpm, removes the EP pipe of 0.5ml, and by 1.5ml
EP be in control liquid and all carry out agarose gel electrophoresis；
2, agarose gel electrophoresis:
(1) glue is joined: according to genomic fragment size, the agarose gel of configuration respective concentration.Take appropriate agar and TAE
Buffer, in conical flask, shakes up and seals with tinfoil, put into microwave oven and boil, and well-done gel answers bubble-free, uniform color.Will
Well-done gel is put in the water-bath of 65 DEG C, when gel is cooled to 65 DEG C, adds appropriate SYBR Green I fluorescence dye
Material, pours into gel in the clean electrophoresis mould installed in time.After gel solidifies completely, careful extracts comb；
(2) electrophoresis prepares: gel is put into electrophoresis tank centre together with glue torr, and the direction in glue hole is towards negative pole, past
Electrophoresis tank is poured a certain amount of TAE buffer to just being flooded by gel into；
(3) loading: sample, 6 × bromophenol blue are mixed in the ratio of 5:1, according to the order loading of regulation after gently getting rid of；
(4) electrophoresis: after loading, covers electrophoresis tank lid, connects electrophresis apparatus and electrophoresis tank, arranges electrophoretic parameters: 120V
30min；
3, DNA reclaims:
(1) chopping of the blob of viscose containing target DNA that will scale off, puts into and is drilled hole in bottom and puts into fritter qualitative filter paper
0.5ml EP pipe in, then the EP pipe of this 0.5ml is inserted in the EP pipe of a 1.5ml；
(2) the EP pipe packaged is centrifuged 5～10min under 4 DEG C of 10000rpm, removes the EP pipe of 0.5ml, and by 1.5ml
EP be in control liquid liquid-transfering gun constant volume；
(3) 3mol/L NaAc (pH 5.5) and the dehydrated alcohol of 2 times of volumes of 1/10 volume are added ,-20 DEG C of freezings
More than 30min.Precipitation is dissolved in 20 μ l TE solution (pH 7.8) after drying.
4, SYBR Green I fluorescence staining:
(1) (dilution ratio is according to SYBR to add the SYBR Green I working solution good with the DNA redissolution isopyknic dilution of liquid
The description of Green I), mixing；
(2) place 30 minutes.
5, detection and interpretation of result:
(1) DNA solution dyeed is placed on ultraviolet spectrophotometer, at 260nm wavelength, reads fluorescent value respectively
A260；
(2) according to WdsDNATarget DNA content in the μ g/ml formula scales testing sample of=A260 × 50.
With reference to Fig. 1, the application present invention detects the DNA content of genetic fragment length about 1kb in testing sample A, concurrently sets
The standard control of one group of known dna concentration (30ng/ μ l), obtains electrophoretogram 1, then warp after coarse filtration and agarose gel electrophoresis
Cross DNA reclaim, SYBR Green I fluorescence staining and detection and interpretation of result, record result such as table 2:
Table 2
As in figure 2 it is shown, this EP pipe comprises EP 21 and the qualitative filter paper 22 of the hole-drilled 0.5ml in bottom.Such as Fig. 3 institute
Showing, this EP pipe comprises and is drilled hole in bottom and puts into the EP pipe 32 of EP pipe 31 and 1.5ml of 0.5ml of fritter qualitative filter paper.
As shown in Figure 4, this test kit include shown in Fig. 2 be drilled in bottom hole and put into fritter qualitative filter paper 0.5ml EP pipe
Being inserted in shown in 41 and Fig. 3 is drilled hole and puts into the EP pipe 42 of 1.5ml of 0.5ml EP of fritter qualitative filter paper in bottom.
Above in conjunction with accompanying drawing, the present invention is exemplarily described, it is clear that the present invention implements not by aforesaid way
Restriction, as long as have employed method design and the various improvement that carry out of technical scheme of the present invention, or the most improved direct application
In other occasion, all within protection scope of the present invention.
Claims (10)
1. the SYBR Green I immue quantitative detection reagent box of a detection by quantitative DNA content, it is characterised in that include box body,
Frame, an EP pipe and the 2nd EP pipe, wherein,
In described box body, support is set；
Described support is provided with 4-6 pipe support；
A described EP pipe is arranged on the first pipe support, is provided with boring, is provided with qualitative filter paper in pipe bottom a described EP pipe；
Described 2nd EP pipe is arranged on the second pipe support, is provided with boring, is provided with qualitative filter paper in pipe bottom described 2nd EP pipe.
2. the SYBR Green I immue quantitative detection reagent box of detection by quantitative DNA content as claimed in claim 1, its feature exists
In, also include that lid, described lid side are connected to box body and for fastening box body.
3. the SYBR Green I immue quantitative detection reagent box of the detection by quantitative DNA content as described in claim 1 and 2, its feature
Being, a described EP pipe is the EP pipe of 0.5ml or 1.5ml, and described 2nd EP pipe is the EP pipe of 0.5ml or 1.5ml.
4. the detection method of the SYBR Green I immue quantitative detection reagent box of detection by quantitative DNA content as described in claim 1-3,
It is characterized in that, comprise the steps:
(1) coarse filtration；
(2) agarose gel electrophoresis；
(3) DNA reclaims；
(4) SYBR Green I fluorescence staining；
(5) detection；
(6) interpretation of result.
5. the detection method of the SYBR Green I immue quantitative detection reagent box of detection by quantitative DNA content as claimed in claim 4, its
Being characterised by, step (1) including:
(1-1) take testing sample to put in the EP pipe that bottom is drilled hole the 0.5ml that puts into fritter qualitative filter paper, then by this
The EP pipe of individual 0.5ml is inserted in the EP pipe of a 1.5ml；
(1-2) the EP pipe that packages is centrifugal, remove the EP pipe of 0.5ml, and the EP of 1.5ml is in control liquid all carries out agar
Sugar gel electrophoresis.
The detection side of the SYBR Green I immue quantitative detection reagent box of detection by quantitative DNA content the most as claimed in claims 4 and 5
Method, it is characterised in that step (2) including:
(2-1) glue is joined；
(2-2) electrophoresis prepares；
(2-3) loading；
(2-4) electrophoresis.
7. the detection method of the SYBR Green I immue quantitative detection reagent box of detection by quantitative DNA content as described in claim 4-6,
It is characterized in that, step (3) including:
(3-1) chopping of the blob of viscose containing target DNA that will scale off, puts into and is drilled hole in bottom and puts into fritter qualitative filter paper
In the EP pipe of 0.5ml, then the EP pipe of this 0.5ml is inserted in the EP pipe of a 1.5ml；
(3-2) the EP pipe packaged is centrifuged, removes the EP pipe of 0.5ml, and the EP of 1.5ml is in control liquid capacity-fixed；
(3-3) NaAc and dehydrated alcohol are added, freezing；Precipitation is dissolved in TE solution after drying.
8. the detection method of the SYBR Green I immue quantitative detection reagent box of detection by quantitative DNA content as described in claim 4-7,
It is characterized in that, step (4) including:
(4-1) the SYBR Green I working solution good with the DNA redissolution isopyknic dilution of liquid, mixing are added；
(4-2) about 30 minutes are placed.
9. the detection method of the SYBR Green I immue quantitative detection reagent box of detection by quantitative DNA content as described in claim 4-8,
It is characterized in that, step (5) including:
(5-1) DNA solution dyeed is placed on ultraviolet spectrophotometer, at 260nm wavelength, reads fluorescent value respectively
A260；
(5-2) according to WdsDNATarget DNA content in the μ g/ml formula scales testing sample of=A260 × 50.
10. the detection method of the SYBR Green I immue quantitative detection reagent box of detection by quantitative DNA content as claimed in claim 6,
It is characterized in that, (2-1) joins glue: according to genomic fragment size, the agarose gel of configuration respective concentration；Take appropriate agar
With in TAE buffer to conical flask, shaking up and seal with tinfoil, put into microwave oven and boil, well-done gel answers bubble-free, color and luster equal
Even；Well-done gel is put in the water-bath of 65 DEG C, when gel is cooled to 65 DEG C, add appropriate SYBRGreen I glimmering
Photoinitiator dye, pours into gel in the clean electrophoresis mould installed in time；After gel solidifies completely, careful extracts comb；
(2-2) electrophoresis prepares: gel is put into electrophoresis tank centre together with glue torr, and the direction in glue hole is towards negative pole, toward electrophoresis tank
In pour a certain amount of TAE buffer into just being flooded by gel；(2-3) loading: by sample, 6 × bromophenol blue in the ratio of 5:1
Mixing, according to the order loading of regulation after gently getting rid of；(2-4) electrophoresis: after loading, covers electrophoresis tank lid, connects electrophresis apparatus and electrophoresis
Groove, arranges electrophoretic parameters: 120V 30min.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610737798.7A CN106198479A (en) | 2016-08-26 | 2016-08-26 | The SYBR Green I immue quantitative detection reagent box of a kind of detection by quantitative DNA content and detection method thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610737798.7A CN106198479A (en) | 2016-08-26 | 2016-08-26 | The SYBR Green I immue quantitative detection reagent box of a kind of detection by quantitative DNA content and detection method thereof |
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| CN106198479A true CN106198479A (en) | 2016-12-07 |
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1661104A (en) * | 2004-12-17 | 2005-08-31 | 浙江大学 | Signal detection method for gene chip of oligonucleotide with length more than 50 basic groups |
| CN101376883A (en) * | 2007-08-31 | 2009-03-04 | 中国科学院沈阳应用生态研究所 | Simple and efficient PCR product purification recovery method and special equipment thereof |
| CN101831507A (en) * | 2010-05-10 | 2010-09-15 | 中国水产科学研究院长江水产研究所 | Real-time fluorescence quantitative PCR (polymerase chain reaction) detection method of grass carp reovirus SYBR Green I |
| CN102174653A (en) * | 2011-03-01 | 2011-09-07 | 广西壮族自治区兽医研究所 | Haemophilus parasuis real-time fluorescent quantitative PCR (polymerase chain reaction) detection method |
| CN204142680U (en) * | 2014-07-31 | 2015-02-04 | 北京海思特临床检验所有限公司 | Single cell gel electrophoresis is adopted to detect the kit of DNA chain fracture |
| CN205473772U (en) * | 2015-11-13 | 2016-08-17 | 山东世通检测评价技术服务有限公司 | Multiple PCR detect reagent box of donkey -hide gelatin true and false |
| CN206132619U (en) * | 2016-08-26 | 2017-04-26 | 周辉 | Quantitative determination kit |
-
2016
- 2016-08-26 CN CN201610737798.7A patent/CN106198479A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1661104A (en) * | 2004-12-17 | 2005-08-31 | 浙江大学 | Signal detection method for gene chip of oligonucleotide with length more than 50 basic groups |
| CN101376883A (en) * | 2007-08-31 | 2009-03-04 | 中国科学院沈阳应用生态研究所 | Simple and efficient PCR product purification recovery method and special equipment thereof |
| CN101831507A (en) * | 2010-05-10 | 2010-09-15 | 中国水产科学研究院长江水产研究所 | Real-time fluorescence quantitative PCR (polymerase chain reaction) detection method of grass carp reovirus SYBR Green I |
| CN102174653A (en) * | 2011-03-01 | 2011-09-07 | 广西壮族自治区兽医研究所 | Haemophilus parasuis real-time fluorescent quantitative PCR (polymerase chain reaction) detection method |
| CN204142680U (en) * | 2014-07-31 | 2015-02-04 | 北京海思特临床检验所有限公司 | Single cell gel electrophoresis is adopted to detect the kit of DNA chain fracture |
| CN205473772U (en) * | 2015-11-13 | 2016-08-17 | 山东世通检测评价技术服务有限公司 | Multiple PCR detect reagent box of donkey -hide gelatin true and false |
| CN206132619U (en) * | 2016-08-26 | 2017-04-26 | 周辉 | Quantitative determination kit |
Non-Patent Citations (3)
| Title |
|---|
| L螺S丝D钉: "琼脂糖凝胶电泳操作标准流程"的文章", 《百度文库》 * |
| 刘歆等: "基于SYBRGreenI的双链DNA定量方法", 《中国生物工程杂志》 * |
| 李凌主编: "《生物化学与分子生物学实验指导》", 31 August 2015, 人民军医出版社 * |
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