A kind of kit and its application method extracting pathogenic nucleic acid from animal tissue sampleTechnical field
The present invention relates to field of biotechnology, specially a kind of kit for extracting pathogenic nucleic acid from animal tissue sample andIts application method.
Background technique
In recent years, the technology of molecular biology is mainly based on to the detection of animal epidemic, and the base of molecular biologyPlinth is nucleic acid extraction.Wherein phenol extraction method, isopropanol precipitating method, formamide cracking process and TriZol method are the classical nucleic acid of comparisonMethod for extracting, improvements of many methods is carried out on the basis of these methods at present, before three kinds of methods be utilizationProteinase K and lauryl sodium sulfate (SDS) digest smudge cells, and TriZol method is broken thin with guanidinium isothiocyanate digestionBorn of the same parents.After clasmatosis formamide method be using high concentration formamide depolymerizing protein matter and DNA combination, then using dialyse comeDNA sample is handled, but other three kinds of methods are then first to remove isolating protein with phenol/chloroform, then use ethyl alcohol or isopropanol precipitating respectivelyDNA。
Currently, extracting the reagent that pathogenic nucleic acid from animal tissue sample uses, saved when stored in refrigerating chamber, thanIf -20 DEG C of guanidinium isothiocyanate save, in test, need to take out from refrigerating chamber, if influencing the work of reagent without cryo-conservationProperty or failure, thus influence test as a result, causing test result inaccurate.In consideration of it, we provide a kind of extraction animal groupsThe kit and its application method of etiology nucleic acid in tissue samples.
Summary of the invention
The purpose of the present invention is to provide a kind of kit for extracting pathogenic nucleic acid from animal tissue sample and its usersMethod, if to solve to propose that nowadays reagent is without cryo-conservation, vulnerable problem in above-mentioned background technique.
To achieve the above object, the invention provides the following technical scheme:
A kind of kit and its application method extracting pathogenic nucleic acid from animal tissue sample, including reagent box body and placementIn several test tubes of the kit body interior, the reagent box body uses the top square hollow for open interiorStructure, the top side edge of the reagent box body are connected by a hinge and have box cover, the reagent box body and box cover it is interiorWall is provided with insulating layer, is embedded at the top of the reagent box body and is equipped with test tube fixed plate, opens in the test tube fixed plateTest tube hole equipped with several uniform equidistant arrangements, the intrinsic bottom of kit are equipped with test tube firm banking, the examinationOffer that, position identical as the test tube hole number be corresponding and the compatible test tube slot of size at the top of pipe firm banking,The inner sidewall of the test tube firm banking and bottom are hollow structure, and inner sidewall and bottom are filled with anti-freeze cooling liquid, instituteTest tube firm banking is stated closely to weld with the reagent box body；
Bottom is equipped with cooling device in the reagent box body, the cooling device include evaporator, compressor, condenser andFilter, the evaporator are set in the test tube firm banking and are immersed in anti-freeze cooling liquid, the evaporator, compressionMachine, condenser and filter are sequentially connected in series by pipeline, and the compressor, condenser and filter are installed on the test tubeThe outside of firm banking is simultaneously fixed in kit inner body wall.
Preferably, the insulating layer uses inorganic heat insulation material, including aerogel blanket, mineral wool or rock wool.
Preferably, the reagent box body symmetrically offers fixinig plate slot, Mei Gesuo far from a lateral roof of hingeIt states and offers fixing threaded hole at the center of fixinig plate slot, the box cover is symmetrically arranged with fixation far from one end two sides of hingePiece, the fixinig plate is corresponding with the position of the fixinig plate slot, and fixinig plate and fixinig plate slot are mating, Mei GesuoIt states and is mounted on screw thread bolt at the center of fixinig plate, the screw thread bolt is threadedly coupled with the fixing threaded hole.
Preferably, the test tube hole is provided with internal screw thread wall, it is provided on the test tube and internal screw thread wall positionCorresponding external screw thread wall, the length of the test tube are greater than the height of the reagent box body, in the top exterior walls of the test tubeIt is provided with reagent label.
Preferably, the bottom surface of the box cover offers the compatible sealing of size corresponding with the test tube hole siteSlot, the sum of the depth of the seal groove and height of reagent box body are adapted with the length dimension of test tube.
Preferably, the evaporator is metal steel structure, the evaporator is provided with back-shaped tube passage；The back-shaped pipeThe input end in channel connects filter by pipeline, and the outlet end of the back-shaped tube passage passes through pipeline connect compressor.
Preferably, the side length of the reagent box body be 15-25cm, the test tube fixed plate with a thickness of 3-5cm, instituteStating test tube side length is 17-28cm, the box cover with a thickness of 4-8cm, the depth of the seal groove is 2-5cm.
Preferably, being equipped with handle in the middle part of the top surface of the box cover.
Preferably, its specifically used method is as follows:
S1: by rotating screw thread bolt, box cover is opened, takes out all test tubes in reagent box body, and all test tubes are moved toSterilizing, sterilizing time 10-15min are carried out in sterilization box；
S2: the reagent for extracting pathogenic nucleic acid from animal tissue sample is respectively put into test tube hole, and the amount of reagent of each test tube is notMore than 2/3rds of test tube capacity；
S3: according to the reagent type in each test tube, reagent label is bonded on the outer wall of test tube；
S4: the test tube equipped with reagent is sequentially placed into reagent box body, and the bottom of test tube is made to be inserted into the fixed bottom of test tubeIn the test tube slot of seat, and test tube is rotated, is threadedly coupled external screw thread wall with internal screw thread wall；
S5: box cover is closed, and corresponds the seal groove of box cover and test tube, and matching by screw thread bolt and fixing threaded holeIt closes, box cover is fixed with reagent box body；
S6: evaporator, compressor, condenser and filter are turned on external power supply, make its normal work, to anti-freeze cooling liquidCool down, chilled storage is carried out to test tube.
Compared with prior art, the beneficial effects of the present invention are:
1, the kit of this extraction pathogenic nucleic acid from animal tissue sample and its application method in reagent box body by installingCooling device, each electronic component after powered up, cool down to test tube firm banking, to carry out to the reagent in test tube lowTemperature saves, and prevents reagent from causing to fail because of temperature raising.
2, the kit of this extraction pathogenic nucleic acid from animal tissue sample and its application method pass through outside the top of test tubeReagent label is set on wall, and personnel convenient to use look for corresponding reagent in test, while preventing from malfunctioning, setting in evaporatorBack-shaped tube passage, increase coolant in evaporator heat exchange rate, so that the chilling rate of the present embodiment is faster, and save energySource.
Detailed description of the invention
Fig. 1 is the configuration schematic diagram of reagent box body of the present invention；
Fig. 2 is the structural schematic diagram after test tube of the present invention insertion；
Fig. 3 is the structural schematic diagram of reagent box body of the present invention；
Fig. 4 is the cross section structure schematic diagram of reagent box body of the present invention；
Fig. 5 is the overlooking structure figure of test tube firm banking of the present invention；
Fig. 6 is the structure connection figure of cooling device of the present invention.
In figure: 1, reagent box body；2, test tube fixed plate；21, test tube hole；22, internal screw thread wall；3, fixinig plate slot；4,Fixing threaded hole；5, box cover；51, handle；6, seal groove；7, fixinig plate；8, screw thread bolt；9, test tube；91, reagent label；92, outerThreaded walls；10, test tube firm banking；101, test tube slot；102, anti-freeze cooling liquid；103, cooling device；1031, evaporator；1032, back-shaped tube passage；1033, compressor；1034, condenser；1035, filter；11, insulating layer.
Specific embodiment
Below in conjunction with the embodiment of the present invention, technical scheme in the embodiment of the invention is clearly and completely described,Obviously, described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Based in the present inventionEmbodiment, every other embodiment obtained by those of ordinary skill in the art without making creative efforts, allBelong to the scope of protection of the invention.
Embodiment 1
A kind of kit extracting pathogenic nucleic acid from animal tissue sample, as shown in Figure 1, including reagent box body 1 and placementSeveral test tubes 9 inside reagent box body 1, the cube structure that reagent box body 1 uses top hollow for open interior,The top side edge of reagent box body 1, which is connected by a hinge, box cover 5, and the inner wall of reagent box body 1 and box cover 5 is provided withInsulating layer 11, the top of reagent box body 1, which is embedded, is equipped with test tube fixed plate 2, offered in test tube fixed plate 2 it is several uniformly etc.Test tube hole 21 away from arrangement, the bottom in reagent box body 1 are equipped with test tube firm banking 10, the top of test tube firm banking 10Offer that, position identical as 21 quantity of test tube hole be corresponding and the compatible test tube slot 101 of size, test tube firm banking 10Inner sidewall and bottom are hollow structure, and inner sidewall and bottom are filled with anti-freeze cooling liquid 102, test tube firm banking 10 and examinationAgent box body 1 closely welds；
Further, as shown in Figure 4-Figure 6, bottom is equipped with cooling device 103 in reagent box body 1, and cooling device 103 includesEvaporator 1031, compressor 1033, condenser 1034 and filter 1035, evaporator 1031 are set in test tube firm banking 10And be immersed in anti-freeze cooling liquid 102, evaporator 1031, compressor 1033, condenser 1034 and filter 1035 pass through pipelineIt is sequentially connected in series, compressor 1033, condenser 1034 and filter 1035 are installed on the outside of test tube firm banking 10 and consolidateDue on 1 inner wall of reagent box body.
Specifically, insulating layer 11 uses inorganic heat insulation material, including aerogel blanket, mineral wool or rock wool, there is certain protectThe material of temp effect can reach A grades of fire prevention, and as the preferred of the present embodiment, insulating layer 11 uses mineral wool material, have atType is good, bulk density is small, thermal conductivity is low, heat preservation is insulated, sound absorption qualities are good, corrosion-resistant, stable chemical performance.
By installing cooling device 103 in reagent box body 1 in the present embodiment, each electronic component is after powered up, rightTest tube firm banking 10 cools down, to carry out cryo-conservation to the reagent in test tube 9, prevents reagent from causing because of temperature raisingFailure.
Embodiment 2
As second of embodiment of the invention, as shown in Fig. 2, reagent box body 1 is symmetrically opened up far from a lateral roof of hingeThere is fixinig plate slot 3, fixing threaded hole 4, one end two sides of the box cover 5 far from hinge are offered at the center of each fixinig plate slot 3It is symmetrically arranged with fixinig plate 7, fixinig plate 7 is corresponding with the position of fixinig plate slot 3, and fixinig plate 7 and 3 grafting of fixinig plate slotCooperate, is mounted on screw thread bolt 8 at the center of each fixinig plate 7, screw thread bolt 8 is threadedly coupled with fixing threaded hole 4.
Further, test tube hole 21 is provided with internal screw thread wall 22, is provided on test tube 9 corresponding with 22 position of internal screw thread wallExternal screw thread wall 92 so that fixed stronger of test tube 9, the length of test tube 9 is greater than the height of reagent box body 1, test tube 9Reagent label 91 is provided in top exterior walls, personnel convenient to use look for corresponding reagent in test, while preventing from malfunctioning.
Specifically, the bottom surface of box cover 5 offers the compatible seal groove 6 of size corresponding with 21 position of test tube hole, sealingThe sum of the depth of slot 6 and height of reagent box body 1 are adapted with the length dimension of test tube 9, it is worth noting that, seal groove 6Interior the phenomenon that being made using rubber material, prevent leakage.
It is worth noting that evaporator 1031 is metal steel structure, back-shaped tube passage 1032 is provided in evaporator 1031；The input end of back-shaped tube passage 1032 connects filter 1035 by pipeline, and the outlet end of back-shaped tube passage is connected by pipeline pressesContracting machine 1033, the interior back-shaped tube passage 1032 being arranged of evaporator 1031, increases coolant in 1031 heat transfer rate of evaporator, so thatThe chilling rate of the present embodiment is faster, and energy saving.
In addition, the side length of reagent box body 1 be 15-25cm, test tube fixed plate 2 with a thickness of 3-5cm, 9 side length of test tubeFor 17-28cm, box cover 5 with a thickness of 4-8cm, the depth of seal groove 6 is 2-5cm, as the preferred of the present embodiment, kit sheetThe side length of body 1 is 20cm, test tube fixed plate 2 with a thickness of 5cm, 9 side length of test tube is 25cm, box cover 5 with a thickness of 6cm, it is closeThe depth of sealing groove 6 is 5cm.
It is worth noting that, as shown in figure 3, box cover 5 top surface in the middle part of handle 51 is installed, facilitate and carried.
The kit of extraction pathogenic nucleic acid from animal tissue sample of the invention, specifically used method are as follows:
S1: by rotating screw thread bolt 8, box cover 5 is opened, takes out all test tubes 9 in reagent box body 1, and by all test tubes 9It moves to and carries out sterilizing in sterilization box, sterilizing time 10-15min；
S2: the reagent for extracting pathogenic nucleic acid from animal tissue sample is respectively put into test tube hole 21, the amount of reagent of each test tube 9No more than 2/3rds of test tube capacity；
S3: according to the reagent type in each test tube 9, reagent label 91 is bonded on the outer wall of test tube 9；
S4: the test tube 9 equipped with reagent is sequentially placed into reagent box body 1, and so that the bottom of test tube 9 is inserted into test tube and fixesIn the test tube slot 101 of pedestal 10, and test tube 9 is rotated, is threadedly coupled external screw thread wall 92 with internal screw thread wall 22；
S5: box cover 5 is closed, and corresponds the seal groove 6 of box cover 5 and test tube 9, and passes through screw thread bolt 8 and fixed spiral shellBox cover 5 is fixed with reagent box body 1 in the cooperation in hole 4；
S6: being turned on external power supply for evaporator 1031, compressor 1033, condenser 1034 and filter 1035, keeps it normalWork, cools down to anti-freeze cooling liquid 102, carries out chilled storage to test tube 9.
The basic principles, main features and advantages of the present invention have been shown and described above.The technology of the industryFor personnel it should be appreciated that the present invention is not limited to the above embodiments, described in the above embodiment and specification is only the present inventionPreference, be not intended to limit the invention, without departing from the spirit and scope of the present invention, the present invention also has variousChanges and improvements, these changes and improvements all fall within the protetion scope of the claimed invention.The claimed scope of the invention is by instituteAttached claims and its equivalent thereof.