CN107760602A - A kind of cartridge for PCR primer purifying - Google Patents

A kind of cartridge for PCR primer purifying Download PDF

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Publication number
CN107760602A
CN107760602A CN201711080542.4A CN201711080542A CN107760602A CN 107760602 A CN107760602 A CN 107760602A CN 201711080542 A CN201711080542 A CN 201711080542A CN 107760602 A CN107760602 A CN 107760602A
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CN
China
Prior art keywords
magnetosheath
metal bar
pcr primer
cartridge
hole
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN201711080542.4A
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Chinese (zh)
Inventor
王炜
李智洋
翁国武
曲文静
童长华
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Nanjing Colbert Medical Technology Co Ltd
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Nanjing Colbert Medical Technology Co Ltd
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Publication date
Application filed by Nanjing Colbert Medical Technology Co Ltd filed Critical Nanjing Colbert Medical Technology Co Ltd
Priority to CN201711080542.4A priority Critical patent/CN107760602A/en
Publication of CN107760602A publication Critical patent/CN107760602A/en
Pending legal-status Critical Current

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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/10Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1003Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
    • C12N15/1006Extracting 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

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  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Plant Pathology (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Centrifugal Separators (AREA)

Abstract

The invention discloses a kind of cartridge for PCR primer purifying, through hole is offered on the upper lid of cartridge, magnetosheath is provided with through hole, circular protrusion is provided with the middle part of magnetosheath, spring is arranged with the magnetosheath of the bottom of circular protrusion, metal bar is provided with magnetosheath, the bottom of metal bar is that diameter is slightly less than the spherical of magnetosheath internal diameter, metal bar upper end is covered on stretching out, and lower end is inserted into the bottom of magnetosheath, and metal bar can drive magnetosheath to move up and down;Several centrifuge tubes are provided with base.Under the magnetic fields in the external world, metal bar can produce magnetic, and after removing external magnetic field, its magnetic can be completely eliminated.Pass through the up and down motion of metal bar and magnetosheath, the crawl and release of magnetic bead can be completed, capture, washing and elution to nucleic acid is realized by shirtsleeve operation can, PCR primer after purification is finally given, avoids influence of the caused aerosol loss to environment in PCR primer purge process.

Description

A kind of cartridge for PCR primer purifying
Technical field
The present invention relates to a kind of biotinylated nucleic acid molecule extraction element, is specially that a kind of PCR primer purge process that is used for prevents The cartridge of Aerosol Pollution.
Background technology
PCR primer typically will all directly affect containing excessive primer, Taq DNA enzymatics and dNTP, the presence of these compositions To processes such as follow-up digestion, double deoxidation PCR sequencing reactions, it is therefore necessary to remove, but because PCR primer copy number is big (generally 1013Copy/ml), significantly larger than PCR detects the limit of copy number, and during purifying, is very easy to production Angry colloidal sol is spilt into environment, and environment is polluted, and the pollution is difficult to eliminate in a short time, solves this problem to molecule Diagnostic work or research are significant.
The content of the invention
The technical problem to be solved in the present invention is also easy to produce lacking for the pollution of aerosol when being and overcoming existing PCR primer to purify Fall into, there is provided a kind of cartridge for PCR primer purifying.
In order to solve the above-mentioned technical problem, the invention provides following technical scheme:
A kind of cartridge for PCR primer purifying, including upper lid, circular housing and the base installed with one heart, it is circular outer Shell and base can rotate relative to upper lid together, offer through hole on the upper lid, magnetosheath is provided with through hole, the magnetosheath Middle part is provided with circular protrusion, and spring is arranged with the magnetosheath of the bottom of circular protrusion, metal bar, metal are provided with magnetosheath The bottom of rod is that diameter is slightly less than the spherical of magnetosheath internal diameter, and metal bar upper end, which is stretched out, to be covered, and lower end is inserted into the bottom of magnetosheath, gold Category rod can drive magnetosheath to move up and down;Several centrifuge tubes are provided with base.
Further, the through hole of the upper lid and the junction of magnetosheath are provided with sealing ring.Sealing ring is using elasticity Encapsulant is made, and magnetosheath prevents hole top loss of the aerosol from upper lid when moving up and down.
Further, the junction of the base and circular housing is provided with sealing gasket, and sealing gasket is close using elasticity Closure material, when circular housing is rotated with base, aerosol loss can be prevented.
Further, metal bar is formed by the intermetallic composite coating of high magnetic permeability, it is preferred that metal bar uses austenitic stainless steel It is process.Under the magnetic fields in the external world, metal bar can produce magnetic, and after removing external magnetic field, its magnetic can disappear completely Remove.
Further, the centrifuge tube is placed in the hole that base opens up, and the hole on base is interference fitted with centrifuge tube.Hole It is fitted close with centrifuge tube, gapless, is effectively reduced pollution.
Further, 6 centrifuge tubes are placed on base.
The present invention operation principle be:Under the magnetic fields in the external world, metal bar can produce magnetic, remove external magnetic field Afterwards, its magnetic can be completely eliminated.By the up and down motion of metal bar and magnetosheath, the crawl and release of magnetic bead can be completed, is led to Capture, washing and elution of the shirtsleeve operation can realization to nucleic acid are crossed, PCR primer after purification is finally given, avoids Influence of the caused aerosol loss to environment in PCR primer purge process.
Brief description of the drawings
Accompanying drawing is used for providing a further understanding of the present invention, and a part for constitution instruction, the reality with the present invention Apply example to be used to explain the present invention together, be not construed as limiting the invention.In the accompanying drawings:
Fig. 1 is the structural representation of the present invention;
Fig. 2 is the structural representation of magnetosheath;
Fig. 3 is the structural representation of upper lid;
Wherein:1st, metal bar, 2, sealing ring, 3, upper lid, 4, magnetosheath, 5, spring, 6 circular housings, 7, sealing gasket, 8, from Heart pipe, 9, base, 10, through hole, 11, circular protrusion.
Embodiment
The preferred embodiments of the present invention are illustrated below in conjunction with accompanying drawing, it will be appreciated that described herein preferred real Apply example to be merely to illustrate and explain the present invention, be not intended to limit the present invention.
Embodiment 1
As shown in figure 1, a kind of cartridge for PCR primer purifying, including upper lid 3, the and of circular housing 6 installed with one heart Base 9, circular housing 6 and base 9 can be rotated relative to upper lid 3 together, and through hole 10 is offered on upper lid 3, is provided with through hole Magnetosheath 4, the middle part of magnetosheath 4 are provided with circular protrusion 11, and spring 5, magnetosheath 4 are arranged with the magnetosheath of the bottom of circular protrusion 11 Metal bar 1 is inside provided with, the bottom of metal bar 1 is slightly less than the spherical of magnetosheath internal diameter for diameter, and upper lid 3 is stretched out in metal bar upper end, Lower end is inserted into the bottom of magnetosheath 4, and metal bar 1 can drive magnetosheath 4 to move up and down;Several centrifuge tubes 8 are provided with base 9.
Upper lid 3, magnetosheath 4, base 9 use hard transparent plastic injection molding, and metal bar 1 uses the metal of high magnetic permeability difficult to understand Family name's body stainless steel is process, and sealing ring 2, sealing gasket 7 are made of elaxtic seal.Metal bar 1 is inserted into the interior of magnetosheath 4 Kong Li, spring 5 are inserted in from the bottom portion of magnetosheath 4, are then positioned over magnetosheath 4 in the through hole 10 of upper lid 3, the upper ends of through hole 10 are close Seal 2, then upper lid 3 is put into the top of circular housing 6.
6 Standard centrifugal pipes are placed in the hole of base 9,300 μ L lysates, 50 μ L knots are added in the 1st centrifuge tube The particle diameter for closing liquid and 1mg is 500 μm of magnetic nanoparticles, and 500 μ L cleaning solution is separately added into the 2nd~5 centrifuge tube, the 100 μ L eluents are added in 6 centrifuge tubes;200 μ L to be purified PCR primer to be purified is added in the 1st centrifuge tube;It Sealing gasket 7 is inserted in the upper end of base 9 afterwards, then circular housing 6 is installed to the upper end of base 9, completes the assembling of whole cartridge.
Specific extraction operating process is as follows:
(1) circular housing is rotated, by the 1st centrifuge tube on magnetosheath face base, then quickly presses metal bar up and down, Metal bar allows liquid in pipe fully to mix with the magnetosheath quickly liquid internal vibration in the 1st centrifuge tube up and down, allow magnetic bead with Purpose nucleic acid in PCR primer combines;
(2) after to be combined, metal bar top is put into annular magnet, metal bar is magnetized, slowly presses metal bar downwards Dynamic, due to the effect of magnetic force, the magnetic bead in the 1st centrifuge tube is adsorbed to the bottom of magnetosheath, then slowly by metal bar and Magnetosheath moves upwardly together, and magnetic bead leaves the 1st centrifuge tube together with magnetosheath;
(3) circular housing is rotated, magnetosheath is directed at the 2nd centrifuge tube, then inserted metal bar and magnetosheath slowly together Into the cleaning solution of the 2nd centrifuge tube, remain stationary as, remove circular magnet, allow metal bar to lose magnetism, then by quickly repeatedly Move up and down, magnetic bead is separated from magnetosheath, magnetosheath continues to pump, and magnetic bead is mixed in cleaning solution, removes magnetic Other impurity that bead surface combines;
(4) action of 2~3 steps is repeated, magnetic bead is repeated in the 3rd~6 centrifuge tube successively;
(5) repetitive operation (2), the magnetic bead for having washed away nucleic acid is picked up, is then transferred in the 1st hole and abandons.
Extraction process is completed, and the nucleic acid after purification of PCR primer has been obtained in the 6th centrifuge tube.
Finally it should be noted that:The preferred embodiments of the present invention are the foregoing is only, are not intended to limit the invention, Although the present invention is described in detail with reference to the foregoing embodiments, for those skilled in the art, it still may be used To be modified to the technical scheme described in foregoing embodiments, or equivalent substitution is carried out to which part technical characteristic. Within the spirit and principles of the invention, any modification, equivalent substitution and improvements made etc., it should be included in the present invention's Within protection domain.

Claims (6)

1. a kind of cartridge for PCR primer purifying, including upper lid, circular housing and the base installed with one heart, circular housing It can be rotated together relative to upper lid with base, it is characterised in that through hole is offered on the upper lid, magnetosheath is provided with through hole, Circular protrusion is provided with the middle part of the magnetosheath, spring is arranged with the magnetosheath of the bottom of circular protrusion, is provided with magnetosheath Metal bar, the bottom of metal bar are slightly less than the spherical of magnetosheath internal diameter for diameter, and metal bar upper end, which is stretched out, to be covered, and lower end is inserted into magnetic The bottom of set, metal bar can drive magnetosheath to move up and down;Several centrifuge tubes are provided with base.
2. it is used for the cartridge of PCR primer purifying as claimed in claim 1, it is characterised in that the knot of the through hole and magnetosheath Sealing ring is provided with conjunction.
3. it is used for the cartridge of PCR primer purifying as claimed in claim 1, it is characterised in that the base and circular housing Junction be provided with sealing gasket.
4. it is used for the cartridge of PCR primer purifying as claimed in claim 1, it is characterised in that described metal bar is using difficult to understand Family name's body stainless steel is process.
5. it is used for the cartridge of PCR primer purifying as claimed in claim 1, it is characterised in that the centrifuge tube is placed on bottom In the hole that seat opens up, the hole on base is interference fitted with centrifuge tube.
6. it is used for the cartridge of PCR primer purifying as claimed in claim 5, it is characterised in that 6 centrifugations are placed on base Pipe.
CN201711080542.4A 2017-11-06 2017-11-06 A kind of cartridge for PCR primer purifying Pending CN107760602A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201711080542.4A CN107760602A (en) 2017-11-06 2017-11-06 A kind of cartridge for PCR primer purifying

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201711080542.4A CN107760602A (en) 2017-11-06 2017-11-06 A kind of cartridge for PCR primer purifying

Publications (1)

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CN107760602A true CN107760602A (en) 2018-03-06

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111378574A (en) * 2020-03-25 2020-07-07 苏州新海生物科技股份有限公司 Nucleic acid extraction and amplification device
WO2024098324A1 (en) * 2022-11-10 2024-05-16 深圳先进技术研究院 Device and method for purifying nucleic acid and preparing detection reaction liquid

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104059848A (en) * 2014-06-26 2014-09-24 广州冠科生物科技有限公司 Uniform mixing device for nucleic acid extraction
CN107034134A (en) * 2017-06-08 2017-08-11 湖南工业大学 A kind of rotary detection of nucleic acids cartridge
CN107287194A (en) * 2017-07-31 2017-10-24 南京中科拜尔医学技术有限公司 Extraction element
CN207619414U (en) * 2017-11-06 2018-07-17 南京中科拜尔医学技术有限公司 A kind of cartridge for PCR product purifying

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104059848A (en) * 2014-06-26 2014-09-24 广州冠科生物科技有限公司 Uniform mixing device for nucleic acid extraction
CN107034134A (en) * 2017-06-08 2017-08-11 湖南工业大学 A kind of rotary detection of nucleic acids cartridge
CN107287194A (en) * 2017-07-31 2017-10-24 南京中科拜尔医学技术有限公司 Extraction element
CN207619414U (en) * 2017-11-06 2018-07-17 南京中科拜尔医学技术有限公司 A kind of cartridge for PCR product purifying

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111378574A (en) * 2020-03-25 2020-07-07 苏州新海生物科技股份有限公司 Nucleic acid extraction and amplification device
CN111378574B (en) * 2020-03-25 2023-10-27 苏州新海生物科技股份有限公司 Nucleic acid extraction and amplification device
WO2024098324A1 (en) * 2022-11-10 2024-05-16 深圳先进技术研究院 Device and method for purifying nucleic acid and preparing detection reaction liquid

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Application publication date: 20180306

RJ01 Rejection of invention patent application after publication