CN107254468A - A kind of peanut leaf tissue grinding methods and device for being used to extract DNA - Google Patents
A kind of peanut leaf tissue grinding methods and device for being used to extract DNA Download PDFInfo
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- CN107254468A CN107254468A CN201710686110.1A CN201710686110A CN107254468A CN 107254468 A CN107254468 A CN 107254468A CN 201710686110 A CN201710686110 A CN 201710686110A CN 107254468 A CN107254468 A CN 107254468A
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- 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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Abstract
The invention discloses a kind of peanut leaf tissue grinding methods and device for being used to extract DNA, belong to biotechnology technical field, the tip of liquid-transfering gun pipette tips is fired into by ball by alcolhol burner heating, then the liquid-transfering gun pipette tips for being fired into ball using tip are engaged with existing centrifuge tube, peanut leaf tissue mashing after being cooled down through liquid nitrogen, to realize the DNA in the direct peanut leaf tissue extracted in the centrifuge tube after smashing to pieces, relative to smashing to pieces for the peanut leaf tissue realized in prior art by the way of mortar and alms bowl pestle cooperate, alleviate waste of the process of lapping to peanut leaf tissue, it is more suitable for being used to extract peanut leaf tissue DNA when peanut leaf tissue is less；Meanwhile, relative to instrument is ground using automation sample in prior art, reduce peanut leaf DNA extraction cost and saved the usage amount of peanut leaf tissue.
Description
Technical field
The present invention relates to biotechnology technical field, more particularly to a kind of peanut leaf tissue grinder for being used to extract DNA
Method and apparatus.
Background technology
Peanut also known as peanut, belong to annual herb plant, because its oil content is high, are described as " Vegetable meat " by people.
Peanut is in addition to for eating, it may also be used for printing and dyeing, paper industry etc., are the important oil plant of China and industrial crops.China
Peanut cultivation area and production are at the forefront in the world to have very strong competitiveness in the world.But, the current peanut of China
Germ plasm resource it is relatively limited, some disease-resistant degeneration-resistant germ plasm resources are relative to be lacked, and especially aspergillus flavus resisting kind more lacks.
Although conventional crossbreeding, mutation breeding can carry out certain improvement to the gene of peanut, all
It is difficult to realize purposefully to carry out peanut gene improvement.And transgenic breeding can arbitrarily select the main product in production
Plant the importing for carrying out foreign gene, the transfer for the character gene that achieves the goal and the purpose of Genetic improvement.
The common method of existing peanut transgenic breeding has agrobacterium-mediated transformation and particle bombardment.Either use Agrobacterium
Mediated method and particle bombardment are required for being ground peanut leaf tissue, to extract the peanut DNA in peanut leaf tissue.
In prior art, to extract the peanut DNA in peanut leaf tissue, generally cooperated using mortar and alms bowl pestle real in liquid nitrogen
Now to the grinding of peanut leaf tissue, the peanut leaf tissue that grinding is completed, which is transferred to from mortar in corresponding test tube, to be spent
The extraction of raw leaf DNA.
Due to using the grinding of mortar and the mutual cooperation realization of alms bowl pestle to peanut leaf tissue in prior art, largely
Peanut leaf tissue grinder liquid largely stick in mortar and alms bowl pestle, cause the waste of peanut leaf tissue fluid, work as peanut leaf
When organizing less, the method in prior art can not be more used to carry out peanut leaf DNA extraction.How process of lapping pair is mitigated
The waste of peanut leaf tissue, reduction peanut leaf DNA extraction cost has become field technology urgently to be resolved hurrily difficult
Topic.
The content of the invention
The embodiment of the present invention provides a kind of peanut leaf tissue grinding methods and device for being used to extract DNA, it is intended to reduce
To the waste of peanut leaf tissue during peanut leaf tissue grinder, reduction peanut leaf DNA extraction cost.
The concrete technical scheme that the present invention is provided is as follows：
On the one hand, the embodiment of the present invention provides a kind of peanut leaf tissue grinding methods for being used to extract DNA, methods described bag
Include：
Take 0.1 gram ~ 0.4 gram of fresh peanut leaf or take 0.01 gram ~ 0.04 gram of dry peanut blade, be positioned over centrifuge tube
In, wherein, the moisture content of the fresh peanut leaf is more than 40%, and the moisture content of dry peanut blade is less than 20%, the centrifuge tube
The tubulose sample container of the with closure for being 1ml ~ 2ml for volume or gland；
The centrifuge tube of the fresh peanut leaf or the dry peanut blade be will be equipped with as in liquid nitrogen, the liquid nitrogen
Temperature is subzero 196 DEG C；
The tip of 100ul ~ 300ul liquid-transfering gun pipette tips is placed in the flame envelope part of alcolhol burner flame, the liquid-transfering gun pipette tips are treated
Tip is melted as after ball, the liquid-transfering gun pipette tips is removed from the flame envelope part of the alcolhol burner flame, the liquid relief
The tip of rifle pipette tips is through being cooled into ball, and the radius of the ball is 0.1mm ~ 2mm；
Tip is positioned over cooling 3s ~ 10s in liquid nitrogen for the liquid-transfering gun pipette tips of ball；
It is to be installed to there is the fresh flowers leave piece or the centrifuge tube of the dry peanut blade no longer to be sent in the liquid nitrogen
During sound, the centrifuge tube is taken out from liquid nitrogen and is positioned on the centrifugation tube sheet that temperature is subzero 20 DEG C ~ subzero 30 DEG C, and from
The liquid-transfering gun pipette tips are taken out in liquid nitrogen, the liquid-transfering gun pipette tips are inserted in the centrifuge tube, the liquid-transfering gun pipette tips are utilized
The ball at tip quickly smashs the peanut leaf in the centrifuge tube to pieces, to carry out extraction peanut leaf using the centrifuge tube
In tissue the step of DNA.
Optionally, the dry peanut leaf for taking 0.1 gram ~ 0.4 gram of fresh peanut leaf or taking 0.01 gram ~ 0.04 gram
Piece, is positioned in centrifuge tube, is specially：
Take 0.2 gram of fresh peanut leaf or take 0.02 gram of dry peanut blade, be positioned in centrifuge tube, wherein, it is described from
Heart pipe is conical centrifuge tube, and the bottom inner diameter of the conical centrifuge tube is more than the straight of the sophisticated ball of the liquid-transfering gun pipette tips
Footpath, the specification of the conical centrifuge tube is 1.5ml.
Optionally, it is described that tip is positioned over cooling 3s ~ 10s in liquid nitrogen for the liquid-transfering gun pipette tips of ball, be specially：
By inner chamber of the tip insertion tip of tweezers for the liquid-transfering gun pipette tips of ball, with realize the tweezers are stuck in it is described
Serve as the handle of the liquid-transfering gun pipette tips in the inner chamber of liquid-transfering gun pipette tips；
The mobile tweezers, cooling 3s ~ 4s in liquid nitrogen is positioned over by tip for the liquid-transfering gun pipette tips of ball.
Optionally, it is described that tip is positioned over cooling 3s ~ 10s in liquid nitrogen for the liquid-transfering gun pipette tips of ball, be specially：
Handle is inserted into inner chamber of the tip for the liquid-transfering gun pipette tips of ball, the handle is stuck in the liquid-transfering gun to realize
In the inner chamber of pipette tips, wherein, the inner chamber mutual cooperation of the handle and the liquid-transfering gun pipette tips is realized is stuck in institute by the handle
In the inner chamber for stating liquid-transfering gun pipette tips, the handle is one kind in plastic handle, wood handle or metal handle；
The mobile handle, cooling 3s ~ 4s in liquid nitrogen is positioned over by tip for the liquid-transfering gun pipette tips of ball.
Optionally, the tip by 100ul ~ 300ul liquid-transfering gun pipette tips is placed in the flame envelope part of alcolhol burner flame, treats institute
State liquid-transfering gun pipette tips tip melt and turn into ball after, by the liquid-transfering gun pipette tips from the flame envelope part of the alcolhol burner flame
Remove, the tip of the liquid-transfering gun pipette tips is specially through being cooled into ball：
Specification is placed in the flame envelope part of alcolhol burner flame for the tip of 200ul liquid-transfering gun pipette tips, the liquid-transfering gun pipette tips are treated
Tip melt and turn into ball after, the liquid-transfering gun pipette tips are removed from the flame envelope part of the alcolhol burner flame, it is described to move
The tip of liquid rifle pipette tips through being cooled into ball, wherein, the radius of the ball is 1.5mm.
Optionally, it is described to insert the liquid-transfering gun pipette tips in the centrifuge tube, utilize liquid-transfering gun pipette tips tip
Ball quickly smashs the peanut leaf in the centrifuge tube to pieces, is specially：
The liquid-transfering gun pipette tips are inserted in the centrifuge tube, the bottom of the ball and the centrifuge tube at the liquid-transfering gun pipette tips tip
Portion contacts and departed from toward renaturation；
By the extruding force between the ball at liquid-transfering gun pipette tips tip and the bottom of the centrifuge tube, by the centrifuge tube
Peanut leaf tissue mashing, wherein, without visual particulate matter in the peanut leaf tissue after smashing to pieces.
On the other hand, the embodiment of the present invention provides a kind of peanut leaf tissue grinder device for being used to extract DNA, the dress
Put including：
Acquisition module, for taking 0.1 gram ~ 0.4 gram of fresh peanut leaf or taking 0.01 gram ~ 0.04 gram of dry peanut blade,
It is positioned in centrifuge tube, wherein, the moisture content of the fresh peanut leaf is more than 40%, and the moisture content of dry peanut blade is less than
20%, the centrifuge tube is the tubulose sample container of the with closure that volume is 1ml ~ 2ml or gland；
First refrigerating module, for will be equipped with the centrifuge tube of the fresh peanut leaf or the dry peanut blade as
In liquid nitrogen, the temperature of the liquid nitrogen is subzero 196 DEG C；
First processing module, the flame envelope part for the tip of 100ul ~ 300ul liquid-transfering gun pipette tips to be placed in alcolhol burner flame, is treated
After the tip of the liquid-transfering gun pipette tips is melted and turns into ball, by the liquid-transfering gun pipette tips from the flame envelope portion of the alcolhol burner flame
Divide and remove, the tip of the liquid-transfering gun pipette tips is through being cooled into ball, and the radius of the ball is 0.1mm ~ 2mm；
Second refrigerating module, for tip to be positioned over into cooling 3s ~ 10s in liquid nitrogen for the liquid-transfering gun pipette tips of ball；
Second processing module, for the centrifuge tube to be installed for having the fresh flowers leave piece or the dry peanut blade in institute
State when no longer sending sound in liquid nitrogen, the centrifuge tube is taken out from liquid nitrogen be positioned over temperature for subzero 20 DEG C ~ it is subzero 30 DEG C
Centrifugation tube sheet on, and take out from liquid nitrogen the liquid-transfering gun pipette tips, the liquid-transfering gun pipette tips inserted in the centrifuge tube, profit
Quickly the peanut leaf in the centrifuge tube is smashed to pieces with the ball at liquid-transfering gun pipette tips tip, to use the centrifuge tube
The step of carrying out extracting DNA in peanut leaf tissue.
Optionally, the acquisition module specifically for：
Take 0.2 gram of fresh peanut leaf or take 0.02 gram of dry peanut blade, be positioned in centrifuge tube, wherein, it is described from
Heart pipe is conical centrifuge tube, and the bottom inner diameter of the conical centrifuge tube is more than the straight of the sophisticated ball of the liquid-transfering gun pipette tips
Footpath, the specification of the conical centrifuge tube is 1.5ml.
Optionally, second refrigerating module specifically for：
By inner chamber of the tip insertion tip of tweezers for the liquid-transfering gun pipette tips of ball, with realize the tweezers are stuck in it is described
Serve as the handle of the liquid-transfering gun pipette tips in the inner chamber of liquid-transfering gun pipette tips；
The mobile tweezers, cooling 3s ~ 4s in liquid nitrogen is positioned over by tip for the liquid-transfering gun pipette tips of ball.
Optionally, second refrigerating module specifically for：
Handle is inserted into inner chamber of the tip for the liquid-transfering gun pipette tips of ball, the handle is stuck in the liquid-transfering gun to realize
In the inner chamber of pipette tips, wherein, the inner chamber mutual cooperation of the handle and the liquid-transfering gun pipette tips is realized is stuck in institute by the handle
In the inner chamber for stating liquid-transfering gun pipette tips, the handle is one kind in plastic handle, wood handle or metal handle；
The mobile handle, cooling 3s ~ 4s in liquid nitrogen is positioned over by tip for the liquid-transfering gun pipette tips of ball.
Optionally, the first processing module specifically for：
Specification is placed in the flame envelope part of alcolhol burner flame for the tip of 200ul liquid-transfering gun pipette tips, the liquid-transfering gun pipette tips are treated
Tip melt and turn into ball after, the liquid-transfering gun pipette tips are removed from the flame envelope part of the alcolhol burner flame, it is described to move
The tip of liquid rifle pipette tips through being cooled into ball, wherein, the radius of the ball is 1.5mm.
Optionally, the Second processing module specifically for：
The liquid-transfering gun pipette tips are inserted in the centrifuge tube, the bottom of the ball and the centrifuge tube at the liquid-transfering gun pipette tips tip
Portion contacts and departed from toward renaturation；
By the extruding force between the ball at liquid-transfering gun pipette tips tip and the bottom of the centrifuge tube, by the centrifuge tube
Peanut leaf tissue mashing, wherein, without visual particulate matter in the peanut leaf tissue after smashing to pieces.
Beneficial effects of the present invention are as follows：
The embodiment of the present invention provides the peanut tissue grinding methods for extracting DNA, is heated by alcolhol burner by liquid-transfering gun pipette tips
Tip be fired into ball, be then fired into the liquid-transfering gun pipette tips of ball using tip is engaged with existing centrifuge tube, will be through
Peanut leaf tissue mashing after liquid nitrogen cooling, to realize the direct peanut leaf group extracted in the centrifuge tube after smashing to pieces
DNA in knitting, relative to smashing for the peanut leaf tissue realized in prior art by the way of mortar and alms bowl pestle cooperate
It is broken, waste of the process of lapping to peanut leaf tissue is alleviated, is more suitable for being used for extracting peanut when peanut leaf tissue is less
Leaf tissue DNA；Meanwhile, relative to instrument is ground using automation sample in prior art, reduce carrying for peanut leaf DNA
Take cost and save the usage amount of peanut leaf tissue；Due to the method for the embodiment of the present invention, by peanut leaf in centrifuge tube
After piece tissue mashing, it is not necessary to which the peanut leaf tissue after transfer is smashed to pieces can be achieved to carry peanut leaf tissue DNA
Take, it is to avoid the use and cleaning of mortar in prior art, improve the utilization rate of peanut leaf tissue, save simultaneously
The usage amount of liquid nitrogen, it is ensured that the extraction effect of peanut leaf tissue DNA, and centrifuge tube and liquid-transfering gun pipette tips be cost compared with
The common equipment of low exerciser, realizes the quick, inexpensive of peanut leaf tissue and smashs to pieces, save experimentation cost.
Brief description of the drawings
Technical scheme in order to illustrate the embodiments of the present invention more clearly, makes required in being described below to embodiment
Accompanying drawing is briefly described, it should be apparent that, drawings in the following description are only some embodiments of the present invention, for
For those of ordinary skill in the art, on the premise of not paying creative work, other can also be obtained according to these accompanying drawings
Accompanying drawing.
Fig. 1 shows for a kind of method flow of peanut leaf tissue grinding methods for extracting DNA of the embodiment of the present invention
It is intended to；
Fig. 2 melts the structural representation of the liquid-transfering gun pipette tips as ball for the tip used of the embodiment of the present invention；
Fig. 3 melts the liquid-transfering gun pipette tips as ball for the tip of the embodiment of the present invention and is engaged with centrifuge tube for grinding peanut
The schematic diagram of leaf tissue；
Fig. 4 is a kind of structured flowchart for being used to extract DNA peanut leaf tissue grinder device of the embodiment of the present invention.
Embodiment
In order that the object, technical solutions and advantages of the present invention are clearer, below in conjunction with accompanying drawing the present invention is made into
One step it is described in detail, it is clear that described embodiment is only a part of embodiment of the invention, rather than whole implementation
Example.Based on the embodiment in the present invention, what those of ordinary skill in the art were obtained under the premise of creative work is not made
All other embodiment, belongs to the scope of protection of the invention.
Term " first ", " second ", " the 3rd " in description and claims of this specification and above-mentioned accompanying drawing, "
Four ", " 5th ", " the 6th ", " the 7th " and " the 8th " etc.（If there is）It is for distinguishing similar object, without for retouching
State specific order or precedence.It should be appreciated that the data so used can be exchanged in the appropriate case, to retouch here
The embodiments of the invention stated can for example be implemented with the order in addition to those for illustrating or describing herein.
In addition, term " comprising " and " having " and their any deformation, it is intended that covering is non-exclusive to be included, example
Such as, process, method, system, product or the equipment for containing series of steps or unit are not necessarily limited to those clearly listed
Step or unit, but may include not list clearly or for intrinsic other of these processes, method, product or equipment
Step or unit.
For the waste during peanut leaf tissue grinder in reduction prior art to peanut leaf tissue, peanut leaf is reduced
Piece DNA extraction cost, the embodiment of the present invention provides a kind of peanut leaf tissue grinding methods and device for being used to extract DNA,
The tip of liquid-transfering gun pipette tips is fired into by ball by alcolhol burner heating, the liquid-transfering gun pipette tips of ball are then fired into using tip
It is engaged with existing centrifuge tube, the peanut leaf tissue mashing after being cooled down through liquid nitrogen, to realize directly in the centrifuge tube
DNA in the middle peanut leaf tissue extracted after smashing to pieces, relative to what is cooperated in prior art using mortar and alms bowl pestle
Smashing to pieces for the peanut leaf tissue that mode is realized, alleviates waste of the process of lapping to peanut leaf tissue, is more suitable for being used for
Peanut leaf tissue DNA is extracted when peanut leaf tissue is less；Meanwhile, ground relative to automation sample is used in prior art
Instrument, reduces peanut leaf DNA extraction cost and has saved the usage amount of peanut leaf tissue；Due to the embodiment of the present invention
Method, by after peanut leaf tissue mashing in centrifuge tube, it is not necessary to transfer smash to pieces after peanut leaf tissue
Realize extraction to peanut leaf tissue DNA, it is to avoid the use and cleaning of mortar in prior art, improve peanut leaf
The utilization rate of piece tissue, while saving the usage amount of liquid nitrogen, it is ensured that the extraction effect of peanut leaf tissue DNA, Er Qieli
Heart pipe and liquid-transfering gun pipette tips are the common equipment of lower-cost exerciser, realize quick, the low cost of peanut leaf tissue
Smash to pieces, save experimentation cost.
Below in conjunction with the peanut leaf tissue grinder side that is used to extract DNA of the 1 ~ accompanying drawing of accompanying drawing 3 to the embodiment of the present invention
Method is described in detail.
Shown in refer to the attached drawing 1, the peanut leaf tissue grinding methods for being used to extract DNA of the embodiment of the present invention are including as follows
Step：
Step 110：Take 0.1 gram ~ 0.4 gram of fresh peanut leaf or take 0.01 gram ~ 0.04 gram of dry peanut blade, be positioned over
In centrifuge tube.
If the embodiment of the present invention uses fresh peanut leaf, wherein, fresh peanut leaf is from peanut live body
The peanut leaf won, the moisture content of fresh peanut leaf is more than 40%.In the peanut for being used to extract DNA of the embodiment of the present invention
In leaf tissue Ginding process, the weight for the fresh peanut leaf once chosen can be between 0.1 gram ~ 0.4 gram；For once
The specific weight for the fresh peanut leaf chosen, the embodiment of the present invention is not specifically limited, as long as being limited in the embodiment of the present invention
In the range of.
Wherein, it is preferred that the weight for the fresh peanut leaf that the embodiment of the present invention is chosen is 0.1 gram ~ 0.2 gram, so may be used
While effectively to extract the DNA of peanut leaf tissue in the method for ensureing the embodiment of the present invention, it is to avoid to test material peanut
The waste of blade, improves utilization rate and the reduction experimentation cost of peanut leaf.
If the embodiment of the present invention uses dry peanut blade, wherein, dry peanut blade is from the peanut body dried
On the peanut leaf won, the moisture content of dry peanut blade is less than 20%.In the peanut for being used to extract DNA of the embodiment of the present invention
In leaf tissue Ginding process, the weight for the dry peanut blade once chosen can be between 0.01 gram ~ 0.04 gram；For once
The specific weight of the dry peanut blade of selection, the embodiment of the present invention is not specifically limited, as long as limited in the embodiment of the present invention
In the range of.
Wherein, it is preferred that the weight for the dry peanut blade that the embodiment of the present invention is chosen is 0.01 gram ~ 0.02 gram, so may be used
While effectively to extract the DNA of peanut leaf tissue in the method for ensureing the embodiment of the present invention, it is to avoid to test material dried flower
The waste of leave piece, improves utilization rate and the reduction experimentation cost of peanut leaf.
It should be noted that the centrifuge tube that the embodiment of the present invention is used can be the centrifuge tube that volume is 1ml ~ 2ml, its
In, the centrifuge tube is the tubulose sample container of with closure or gland, belongs to the common test equipment in laboratory；Example, this hair
Bright embodiment uses 1ml conical centrifuge tubes or 1.5ml conical centrifuge tubes.
Example, take 0.2 gram of fresh peanut leaf or take 0.02 gram of dry peanut blade, be positioned over 1.5ml point
In the inner chamber 201 of bottom centrifuge tube, and the bottom inner diameter of the conical centrifuge tube is straight more than the sophisticated ball of liquid-transfering gun pipette tips
Footpath.
Step 120：The centrifuge tube of the fresh peanut leaf or the dry peanut blade be will be equipped with as liquid nitrogen
In.
The centrifuge tube that will be equipped with fresh peanut leaf or dry peanut blade is positioned in liquid nitrogen and cooled down, wherein, liquid
The temperature of nitrogen be subzero 196 DEG C, because the temperature of liquid nitrogen is low, it is possible to achieve the temperature of peanut leaf and centrifuge tube rapidly under
Drop, it is to avoid DNA of the peanut leaf in peanut leaf tissue during cooling is destroyed.
Peanut leaf is cooled down using liquid nitrogen, peanut leaf peanut leaf during cooling can be not only avoided
DNA in tissue is destroyed, but also peanut leaf can be carried out after quickly cooling down using liquid nitrogen, can reduce peanut leaf
The grinding difficulty of piece, it is ensured that the integrality for the DNA that process of lapping will not be destroyed in peanut leaf tissue.
Step 130：The tip of 100ul ~ 300ul liquid-transfering gun pipette tips is placed in the flame envelope part of alcolhol burner flame, treats described
The tip of liquid-transfering gun pipette tips is melted as after ball, and the liquid-transfering gun pipette tips are moved from the flame envelope part of the alcolhol burner flame
Go out, the tip of the liquid-transfering gun pipette tips is through being cooled into ball.
Wherein, liquid-transfering gun is one kind of pipettor, be usually used in laboratory it is a small amount of or it is micro liquid pipette, specification is different,
The liquid-transfering gun of different size supports the use different size of pipette tips, and the shape of different manufacturer's productions is also slightly different, but work
Make principle and operating method is basically identical.
The range ability of liquid-transfering gun is usually 0.1ul ~ 5000ul, wherein, the liquid-transfering gun of different range abilities uses difference
Liquid-transfering gun pipette tips, example, the embodiment of the present invention can also may be used using selected range as the pipette tips of 100ul ~ 150ul liquid-transfering gun
Using selected range as the pipette tips of 100ul ~ 200ul liquid-transfering gun, can also selected range for 100ul ~ 300ul liquid-transfering gun rifle
Head.Wherein, it is preferred that the embodiment of the present invention is from liquid-transfering gun pipette tips of the color for the 200ul of yellow, from 200ul liquid relief
Rifle pipette tips are engaged with 1.5ml conical centrifuge tube, and wherein the size and volume of 1.5ml conical centrifuge tube are suitable, and
The intensity and length of 200ul liquid-transfering gun pipette tips are also suitable, and both of which is the conventional instrument in laboratory, and cost compares
It is low, the waste to test material dry peanut blade can be avoided, utilization rate and the reduction experimentation cost of peanut leaf is improved.
As shown in Fig. 2 first, the alcolhol burner that a volume is 60ml or 150ml or 200ml is selected, by alcolhol burner
After lighting, treat that alcolhol burner burnt after a period of time, the tip 302 of 100ul ~ 300ul liquid-transfering gun pipette tips is placed in alcolhol burner
The flame envelope part of flame is heated, after heating a period of time, and the tip 302 of liquid-transfering gun pipette tips is melted, and melts the liquid produced
Body is assembled at the tip 302 of liquid-transfering gun pipette tips.Treat that the liquid that the tip 302 of liquid-transfering gun pipette tips is assembled can form of the invention want
After the ball 303 asked, 100ul ~ 300ul liquid-transfering gun pipette tips are withdrawn to the flame envelope part of alcolhol burner flame, cooled down in atmosphere
Afterwards, at the tip of liquid-transfering gun pipette tips through being cooled into a ball 303, example, as shown in figure 3, in liquid-transfering gun pipette tips
The radius of sophisticated 302 balls 303 through being cooled into is 0.1mm ~ 2mm.
It should be noted that how long just liquid-transfering gun pipette tips are added as alcolhol burner flame envelope for alcolhol burner burning
Heat, the embodiment of the present invention is not specifically limited, and those skilled in the art can be selected according to actual needs.Equally, for
Liquid-transfering gun pipette tips are as the heat time of alcolhol burner flame envelope, and the embodiment of the present invention is also not specifically limited, example, with this area
Technical staff observes the liquid of the tip aggregation of liquid-transfering gun pipette tips, can be at the tip of liquid-transfering gun pipette tips through being cooled into one
Radius is defined for 0.1mm ~ 2mm ball 303.
The tip 302 of liquid-transfering gun pipette tips after alcolhol burner flame envelope heating and melting, after cooling in the point of liquid-transfering gun pipette tips
End forms the ball 303 that a radius is 0.1mm ~ 2mm, wherein, one end of liquid-transfering gun pipette tips of the tip with ball is unlimited
, the other end is closed, wherein, unlimited one end is used for installing tweezers or handle.Formed after being heated through alcolhol burner flame envelope
The liquid-transfering gun pipette tips of ball are sealed, can avoid smashing the peanut leaf in centrifuge tube to pieces with the sophisticated ball of liquid-transfering gun pipette tips
During piece, cause peanut leaf tissue fluid to enter liquid-transfering gun pipette tips, help avoid process of lapping and wave is caused to peanut leaf tissue
Take.
It is preferred that, specification is placed in the outer of alcolhol burner flame by the embodiment of the present invention for the tip of 200ul liquid-transfering gun pipette tips
Flame part, treats that the tip of liquid-transfering gun pipette tips is melted as after ball, by 200ul liquid-transfering gun pipette tips outside alcolhol burner flame
Flame part is removed, the tip of 200ul liquid-transfering gun pipette tips through being cooled into ball, wherein, the radius of the ball is 1.5mm.
Step 140：Tip is positioned over cooling 3s ~ 10s in liquid nitrogen for the liquid-transfering gun pipette tips of ball.
Liquid-transfering gun pipette tips of the tip made in step 130 with ball are positioned over cooling 3s ~ 10s, example in liquid nitrogen
, a handle can be installed in the inner chamber of liquid-transfering gun pipette tips by the big ends 301 of liquid-transfering gun pipette tips, for by step 130
Liquid-transfering gun pipette tips of the tip 302 of middle making with ball 303, which are positioned in liquid nitrogen, to be cooled down.For the shifting that tip is ball
Liquid rifle pipette tips are positioned over the specific cool time in liquid nitrogen, and the embodiment of the present invention is not specifically limited, example, can be by tip
It is positioned in liquid nitrogen for the liquid-transfering gun pipette tips of ball and cools down 5s, it is ensured that tip is reduced to subzero for the liquid-transfering gun pipette tips temperature of ball
Less than 20 DEG C.
Example, handle can be inserted in inner chamber of the tip for the liquid-transfering gun pipette tips of ball, be stuck in handle with realizing
In the inner chamber of liquid-transfering gun pipette tips, the handle of installation is used to put liquid-transfering gun pipette tips of the tip made in step 130 with ball
It is placed in liquid nitrogen and is cooled down；By moving handle, tip is positioned in liquid nitrogen for the liquid-transfering gun pipette tips of ball cool down 3s ~
10s.It is in order to avoid liquid-transfering gun pipette tips are put it should be noted that why install handle in the inner chamber of liquid-transfering gun pipette tips
Being placed in human contact when being cooled down in liquid nitrogen, to liquid nitrogen, can effectively prevent human hand misconnection from contacting liquid nitrogen and human hand is caused to freeze
Wound.Handle is one kind in plastic handle, wood handle or metal handle, example, and the handle can be special handle,
It can be waddy, can also be metallic rod etc., wherein, it is preferred that the handle is wood handle or plastic handle, wood handle
Heat-conducting effect with plastic handle is poor, and user can be avoided to be positioned over liquid-transfering gun pipette tips when being cooled down in liquid nitrogen to user's
Hand causes frostbite.
Example, the tip of tweezers can be inserted in inner chamber of the tip for the liquid-transfering gun pipette tips of ball, to realize tweezer
Subcard serves as the handle of the liquid-transfering gun pipette tips in the inner chamber of liquid-transfering gun pipette tips, for the tip made in step 130 to be carried into circle
The liquid-transfering gun pipette tips of ball, which are positioned in liquid nitrogen, to be cooled down；By mobile tweezers, tip is placed for the liquid-transfering gun pipette tips of ball
3s ~ 4s is cooled down in liquid nitrogen.It should be noted that why the liquid-transfering gun rifle is served as in the inner chamber that tweezers are stuck in into liquid-transfering gun pipette tips
The handle of head, is that human contact, can be effective to liquid nitrogen when being cooled down in order to avoid liquid-transfering gun pipette tips are positioned in liquid nitrogen
Prevent human hand misconnection from contacting liquid nitrogen and causing frostbite to human hand.
Step 150：The centrifuge tube to be installed for having the fresh flowers leave piece or the dry peanut blade is in the liquid
When no longer sending sound in nitrogen, the centrifuge tube is taken out from liquid nitrogen be positioned over temperature for subzero 20 DEG C ~ subzero 30 DEG C from
On heart tube sheet, and the liquid-transfering gun pipette tips are taken out from liquid nitrogen, the liquid-transfering gun pipette tips are inserted in the centrifuge tube, institute is utilized
The ball for stating liquid-transfering gun pipette tips tip quickly smashs the peanut leaf in the centrifuge tube to pieces, to be carried out using the centrifuge tube
The step of extracting DNA in peanut leaf tissue.
Specifically, the centrifuge tube that fresh peanut leaf or dry peanut blade are will be equipped with step 120 is positioned in liquid nitrogen
Cooled down, when the centrifuge tube equipped with fresh flowers leave piece or dry peanut blade no longer sends sound in liquid nitrogen, explanation
The temperature of centrifuge tube equipped with fresh peanut leaf or dry peanut blade falls below the temperature close to liquid nitrogen, i.e., equipped with fresh flowers
When the centrifuge tube of leave piece or dry peanut blade no longer sends sound in liquid nitrogen, illustrate equipped with fresh peanut leaf or dry
The temperature of the centrifuge tube of peanut leaf falls below subzero 196 DEG C.
It should be noted that the centrifuge tube equipped with fresh peanut leaf or dry peanut blade be positioned in liquid nitrogen carry out it is cold
But, its cooling time can not it is oversize can not be too short, such as will be equipped with the centrifugation of fresh peanut leaf or dry peanut blade
Pipe is positioned over the overlong time cooled down in liquid nitrogen, it will the DNA caused because cooling time is long in peanut leaf integrality
It is destroyed；If the centrifuge tube that will be equipped with fresh peanut leaf or dry peanut blade is positioned over the time mistake cooled down in liquid nitrogen
It is short, it will to improve the grinding difficulty of peanut leaf, it is unfavorable for the Ginding process of the peanut leaf tissue using the embodiment of the present invention
Peanut leaf is thoroughly ground to broken, is unfavorable for the extraction of peanut leaf tissue DNA.Present inventor, by substantial amounts of examination
After testing and studying, find when the centrifuge tube equipped with fresh peanut leaf or dry peanut blade be positioned in liquid nitrogen carry out it is cold
But, when the centrifuge tube equipped with fresh flowers leave piece or dry peanut blade no longer sends sound in liquid nitrogen, fresh flowers are shown
The temperature that leave piece or dry peanut blade are cooled down in liquid nitrogen is suitable, you can ensure the integrality of DNA in peanut leaf, drops again
The low grinding difficulty of peanut leaf.
Secondly it is, it is necessary to which explanation, due to different peanut leafs, its moisture content and thermal conductive property are all different, i.e., different
Peanut leaf cooled down in liquid nitrogen required for time it is different, therefore, how to control peanut leaf exactly cold in liquid nitrogen
But correct time, is the technical barrier urgently to be resolved hurrily for perplexing those skilled in the art all the time.Applicant of the present invention
Found after long-term practice and substantial amounts of experiment, the sound sent during by judging that peanut leaf is cooled down in liquid nitrogen,
Can accurately judge that different peanut leafs cool down required correct time in liquid nitrogen, i.e., equipped with fresh peanut leaf or
When the centrifuge tube of dry peanut blade no longer sends sound in liquid nitrogen, show equipped with fresh peanut leaf or dry peanut blade
The temperature of centrifuge tube falls below suitable temperature, is now drawn off being ground, it is ensured that DNA's is complete in peanut leaf
Property, the grinding difficulty of peanut leaf is reduced again.
, will be from when the temperature of the centrifuge tube equipped with fresh flowers leave piece or dry peanut blade falls below subzero 196 DEG C
Heart pipe taken out from liquid nitrogen be positioned over temperature for subzero 20 DEG C ~ it is subzero 30 DEG C between centrifugation tube sheet on, and from liquid nitrogen take out
Liquid-transfering gun pipette tips, rapidly insert liquid-transfering gun pipette tips in the centrifuge tube, quickly will centrifugation using the ball at liquid-transfering gun pipette tips tip
Peanut leaf in pipe is smashed to pieces, after treating that fresh peanut leaf or dry peanut blade in centrifuge tube are smashed to pieces, by liquid-transfering gun
Pipette tips are taken out from centrifuge tube, and the centrifuge tube that then will be equipped with the fresh peanut leaf or dry peanut blade smashed to pieces is transferred to it
On his experimental rig, DNA in the peanut leaf tissue directly extracted using the centrifuge tube in the centrifuge tube is realized.
For extracting peanut leaf tissue using the fresh peanut leaf smashed to pieces in the centrifuge tube or dry peanut blade
Middle DNA detailed process, the embodiment of the present invention is not repeated herein, and those skilled in the art refer to prior art.
It should be noted that centrifugation tube sheet need to be cooled in advance, it is ensured that centrifugation tube sheet temperature be down to subzero 20 DEG C ~
It is subzero between 30 DEG C, during can so ensureing to smash to pieces the fresh peanut leaf or dry peanut blade in centrifuge tube, from
Peanut leaf tissue in heart pipe will not temperature rise it is too many, it is ensured that the DNA in peanut leaf tissue is smash in peanut leaf
During broken, DNA integrality will not be destroyed.
Secondly, it is necessary to be provided with centrifuge tube mounting groove on explanation, centrifugation tube sheet, centrifuge tube can be fixed on to centrifugation
In pipe mounting groove, it is ensured that during using the ball at liquid-transfering gun pipette tips tip is quick, the peanut leaf in centrifuge tube is smashed to pieces,
Centrifuge tube equipped with fresh peanut leaf or dry peanut blade installs firm on centrifugation tube sheet.
Specifically, the temperature of the centrifuge tube of fresh flowers leave piece to be equipped with or dry peanut blade falls below subzero 196 DEG C
When, centrifuge tube is taken out from liquid nitrogen be positioned over temperature for subzero 20 DEG C ~ it is subzero 30 DEG C between centrifugation tube sheet on, then will move
Liquid rifle pipette tips insertion centrifuge tube in, then liquid-transfering gun pipette tips tip ball 303 contacted with the bottom 202 of centrifuge tube toward renaturation and
Depart from；By the extruding force between the ball 303 at liquid-transfering gun pipette tips tip and the bottom 202 of centrifuge tube, by the flower in centrifuge tube
Leave piece tissue mashing, wherein, when the fresh peanut leaf or dry peanut blade in centrifuge tube smash the flower to centrifuge tube to pieces
Without visual particulate matter in raw leaf tissue, you can liquid-transfering gun pipette tips are taken out from centrifuge tube, then will be equipped with smashing to pieces is fresh
The centrifuge tube of peanut leaf or dry peanut blade is transferred on other experimental rigs, is realized and is directly extracted using the centrifuge tube
DNA in peanut leaf tissue in the centrifuge tube.
The embodiment of the present invention provides the peanut tissue grinding methods for extracting DNA, is heated by alcolhol burner by liquid-transfering gun
The tip of pipette tips is fired into ball, and the liquid-transfering gun pipette tips for being then fired into ball using tip are engaged with existing centrifuge tube,
Peanut leaf tissue mashing after being cooled down through liquid nitrogen, to realize the direct peanut leaf extracted in the centrifuge tube after smashing to pieces
DNA in piece tissue, relative to the peanut leaf tissue realized in prior art by the way of mortar and alms bowl pestle cooperate
Smash to pieces, alleviate waste of the process of lapping to peanut leaf tissue, be more suitable for be used for peanut leaf tissue it is less when extract
Peanut leaf tissue DNA；Meanwhile, relative to instrument is ground using automation sample in prior art, reduce peanut leaf DNA
Extraction cost and saved the usage amount of peanut leaf tissue；, will flower in centrifuge tube due to the method for the embodiment of the present invention
After leave piece tissue mashing, it is not necessary to which the peanut leaf tissue after transfer is smashed to pieces can be achieved to peanut leaf tissue DNA
Extraction, it is to avoid the use and cleaning of mortar in prior art, improve the utilization rate of peanut leaf tissue, same to time
Save the usage amount of liquid nitrogen, it is ensured that the extraction effect of peanut leaf tissue DNA, and centrifuge tube and liquid-transfering gun pipette tips are into
This relatively low common equipment of exerciser, realizes the quick, inexpensive of peanut leaf tissue and smashs to pieces, save experimentation cost.
Fig. 4 shows for a kind of structure of peanut leaf tissue grinder device for extracting DNA provided in an embodiment of the present invention
It is intended to.As shown in figure 4, a kind of peanut leaf tissue grinder device for being used to extract DNA provided in an embodiment of the present invention includes：
Acquisition module 401, for taking 0.1 gram ~ 0.4 gram of fresh peanut leaf or taking 0.01 gram ~ 0.04 gram of dry peanut leaf
Piece, is positioned in centrifuge tube, wherein, the moisture content of the fresh peanut leaf is more than 40%, and the moisture content of dry peanut blade is less than
20%, the centrifuge tube is the tubulose sample container of the with closure that volume is 1ml ~ 2ml or gland；
First refrigerating module 402, the centrifuge tube for will be equipped with the fresh peanut leaf or the dry peanut blade
As in liquid nitrogen, the temperature of the liquid nitrogen is subzero 196 DEG C；
First processing module 403, the flame envelope portion for the tip of 100ul ~ 300ul liquid-transfering gun pipette tips to be placed in alcolhol burner flame
Point, after treating that the tip of the liquid-transfering gun pipette tips is melted and turns into ball, by the liquid-transfering gun pipette tips from the alcolhol burner flame
Flame envelope part is removed, and the tip of the liquid-transfering gun pipette tips is through being cooled into ball, and the radius of the ball is 0.1mm ~ 2mm；
Second refrigerating module 404, for tip to be positioned over into cooling 3s ~ 10s in liquid nitrogen for the liquid-transfering gun pipette tips of ball；
Second processing module 405, for the centrifuge tube to be installed for having the fresh flowers leave piece or a dry peanut blade
When no longer sending sound in the liquid nitrogen, the centrifuge tube is taken out from liquid nitrogen be positioned over temperature for subzero 20 DEG C ~ it is subzero
On 30 DEG C of centrifugation tube sheet, and the liquid-transfering gun pipette tips are taken out from liquid nitrogen, the liquid-transfering gun pipette tips are inserted into the centrifuge tube
In, quickly the peanut leaf in the centrifuge tube is smashed to pieces using the ball at liquid-transfering gun pipette tips tip, so as to using described
The step of centrifuge tube carries out extracting DNA in peanut leaf tissue.
Further, acquisition module 401 specifically for：
Take 0.2 gram of fresh peanut leaf or take 0.02 gram of dry peanut blade, be positioned in centrifuge tube, wherein, it is described from
Heart pipe is conical centrifuge tube, and the bottom inner diameter of the conical centrifuge tube is more than the straight of the sophisticated ball of the liquid-transfering gun pipette tips
Footpath, the specification of the conical centrifuge tube is 1.5ml.
Further, the second refrigerating module 404 specifically for：
By inner chamber of the tip insertion tip of tweezers for the liquid-transfering gun pipette tips of ball, with realize the tweezers are stuck in it is described
Serve as the handle of the liquid-transfering gun pipette tips in the inner chamber of liquid-transfering gun pipette tips；
The mobile tweezers, cooling 3s ~ 4s in liquid nitrogen is positioned over by tip for the liquid-transfering gun pipette tips of ball.
Further, the second refrigerating module 404 specifically for：
Handle is inserted into inner chamber of the tip for the liquid-transfering gun pipette tips of ball, the handle is stuck in the liquid-transfering gun to realize
In the inner chamber of pipette tips, wherein, the inner chamber mutual cooperation of the handle and the liquid-transfering gun pipette tips is realized is stuck in institute by the handle
In the inner chamber for stating liquid-transfering gun pipette tips, the handle is one kind in plastic handle, wood handle or metal handle；
The mobile handle, cooling 3s ~ 4s in liquid nitrogen is positioned over by tip for the liquid-transfering gun pipette tips of ball.
Further, first processing module 403 specifically for：
Specification is placed in the flame envelope part of alcolhol burner flame for the tip of 200ul liquid-transfering gun pipette tips, the liquid-transfering gun pipette tips are treated
Tip melt and turn into ball after, the liquid-transfering gun pipette tips are removed from the flame envelope part of the alcolhol burner flame, it is described to move
The tip of liquid rifle pipette tips through being cooled into ball, wherein, the radius of the ball is 1.5mm.
Further, Second processing module 405 specifically for：
The liquid-transfering gun pipette tips are inserted in the centrifuge tube, the bottom of the ball and the centrifuge tube at the liquid-transfering gun pipette tips tip
Portion contacts and departed from toward renaturation；
By the extruding force between the ball at liquid-transfering gun pipette tips tip and the bottom of the centrifuge tube, by the centrifuge tube
Peanut leaf tissue mashing, wherein, without visual particulate matter in the peanut leaf tissue after smashing to pieces.
It should be noted that：What above-described embodiment was provided is used to extract DNA peanut leaf tissue grinder device, only more than
The division progress of each functional module is stated for example, in practical application, as needed can distribute above-mentioned functions by difference
Functional module complete, i.e., the internal structure of equipment is divided into different functional modules, with complete it is described above whole or
Person's partial function.In addition, being used for of providing of above-described embodiment extracts DNA peanut leaf tissue grinder device and extracted with being used for
DNA peanut leaf tissue grinding methods embodiment belongs to same design, and it implements process and refers to embodiment of the method, here
Repeat no more.
The present invention is with reference to method according to embodiments of the present invention, equipment（System）And the flow of computer program product
Figure and/or block diagram are described.It should be understood that can be by every first-class in computer program instructions implementation process figure and/or block diagram
Journey and/or the flow in square frame and flow chart and/or block diagram and/or the combination of square frame.These computer programs can be provided
Instruct the processor of all-purpose computer, special-purpose computer, Embedded Processor or other programmable data processing devices so that
A stream in flow chart can be achieved by the instruction of the computing device of the computer or other programmable data processing devices
The function of being specified in journey or multiple flows and/or one square frame of block diagram or multiple square frames.
These computer program instructions, which may be alternatively stored in, can guide computer or other programmable data processing devices with spy
Determine in the computer-readable memory that mode works so that the instruction being stored in the computer-readable memory, which is produced, to be included referring to
Make the manufacture of device, the command device realize in one flow of flow chart or multiple flows and/or one square frame of block diagram or
The function of being specified in multiple square frames.
These computer program instructions can be also loaded into computer or other programmable data processing devices so that in meter
Series of operation steps is performed on calculation machine or other programmable devices to produce computer implemented processing, thus in computer or
The instruction performed on other programmable devices, which is provided, to be used to realize the flow or multiple flows and/or block diagram in flow chart
A square frame or multiple square frames in specify function the step of.
, but those skilled in the art once know basic creation although preferred embodiments of the present invention have been described
Property concept, then can make other change and modification to these embodiments.So, appended claims are intended to be construed to include excellent
Select embodiment and fall into having altered and changing for the scope of the invention.
Obviously, those skilled in the art can carry out various changes and modification without departing from this hair to the embodiment of the present invention
The spirit and scope of bright embodiment.So, if these modifications and variations of the embodiment of the present invention belong to the claims in the present invention
And its within the scope of equivalent technologies, then the present invention is also intended to comprising including these changes and deformation.
Claims (10)
1. a kind of peanut leaf tissue grinding methods for being used to extract DNA, it is characterised in that methods described includes：
Take 0.1 gram ~ 0.4 gram of fresh peanut leaf or take 0.01 gram ~ 0.04 gram of dry peanut blade, be positioned over centrifuge tube
In, wherein, the moisture content of the fresh peanut leaf is more than 40%, and the moisture content of dry peanut blade is less than 20%, the centrifuge tube
The tubulose sample container of the with closure for being 1ml ~ 2ml for volume or gland；
The centrifuge tube of the fresh peanut leaf or the dry peanut blade be will be equipped with as in liquid nitrogen, the liquid nitrogen
Temperature is subzero 196 DEG C；
The tip of 100ul ~ 300ul liquid-transfering gun pipette tips is placed in the flame envelope part of alcolhol burner flame, the liquid-transfering gun pipette tips are treated
Tip is melted as after ball, the liquid-transfering gun pipette tips is removed from the flame envelope part of the alcolhol burner flame, the liquid relief
The tip of rifle pipette tips is through being cooled into ball, and the radius of the ball is 0.1mm ~ 2mm；
Tip is positioned over cooling 3s ~ 10s in liquid nitrogen for the liquid-transfering gun pipette tips of ball；
It is to be installed to there is the fresh flowers leave piece or the centrifuge tube of the dry peanut blade no longer to be sent in the liquid nitrogen
During sound, the centrifuge tube is taken out from liquid nitrogen and is positioned on the centrifugation tube sheet that temperature is subzero 20 DEG C ~ subzero 30 DEG C, and from
The liquid-transfering gun pipette tips are taken out in liquid nitrogen, the liquid-transfering gun pipette tips are inserted in the centrifuge tube, the liquid-transfering gun pipette tips are utilized
The ball at tip quickly smashs the peanut leaf in the centrifuge tube to pieces, to carry out extraction peanut leaf using the centrifuge tube
In tissue the step of DNA.
2. according to the method described in claim 1, it is characterised in that it is described take 0.1 gram ~ 0.4 gram fresh peanut leaf or
0.01 gram ~ 0.04 gram of dry peanut blade is taken, is positioned in centrifuge tube, is specially：
Take 0.2 gram of fresh peanut leaf or take 0.02 gram of dry peanut blade, be positioned in centrifuge tube, wherein, it is described from
Heart pipe is conical centrifuge tube, and the bottom inner diameter of the conical centrifuge tube is more than the straight of the sophisticated ball of the liquid-transfering gun pipette tips
Footpath, the specification of the conical centrifuge tube is 1.5ml.
3. method according to claim 1 or 2, it is characterised in that described by the liquid-transfering gun pipette tips of the tip for ball
Cooling 3s ~ 10s in liquid nitrogen is positioned over, is specially：
By inner chamber of the tip insertion tip of tweezers for the liquid-transfering gun pipette tips of ball, with realize the tweezers are stuck in it is described
Serve as the handle of the liquid-transfering gun pipette tips in the inner chamber of liquid-transfering gun pipette tips；
The mobile tweezers, cooling 3s ~ 4s in liquid nitrogen is positioned over by tip for the liquid-transfering gun pipette tips of ball.
4. method according to claim 1 or 2, it is characterised in that described by the liquid-transfering gun pipette tips of the tip for ball
Cooling 3s ~ 10s in liquid nitrogen is positioned over, is specially：
Handle is inserted into inner chamber of the tip for the liquid-transfering gun pipette tips of ball, the handle is stuck in the liquid-transfering gun to realize
In the inner chamber of pipette tips, wherein, the inner chamber mutual cooperation of the handle and the liquid-transfering gun pipette tips is realized is stuck in institute by the handle
In the inner chamber for stating liquid-transfering gun pipette tips, the handle is one kind in plastic handle, wood handle or metal handle；
The mobile handle, cooling 3s ~ 4s in liquid nitrogen is positioned over by tip for the liquid-transfering gun pipette tips of ball.
5. method according to claim 1 or 2, it is characterised in that the tip by 100ul ~ 300ul liquid-transfering gun pipette tips
The flame envelope part of alcolhol burner flame is placed in, treats that the tip of the liquid-transfering gun pipette tips is melted as after ball, by the liquid-transfering gun
Pipette tips are removed from the flame envelope part of the alcolhol burner flame, and the tip of the liquid-transfering gun pipette tips is specially through being cooled into ball：
Specification is placed in the flame envelope part of alcolhol burner flame for the tip of 200ul liquid-transfering gun pipette tips, the liquid-transfering gun pipette tips are treated
Tip melt and turn into ball after, the liquid-transfering gun pipette tips are removed from the flame envelope part of the alcolhol burner flame, it is described to move
The tip of liquid rifle pipette tips through being cooled into ball, wherein, the radius of the ball is 1.5mm.
6. according to the method described in claim 1, it is characterised in that described that the liquid-transfering gun pipette tips are inserted into the centrifuge tube
In, quickly the peanut leaf in the centrifuge tube is smashed to pieces using the ball at liquid-transfering gun pipette tips tip, is specially：
The liquid-transfering gun pipette tips are inserted in the centrifuge tube, the bottom of the ball and the centrifuge tube at the liquid-transfering gun pipette tips tip
Portion contacts and departed from toward renaturation；
By the extruding force between the ball at liquid-transfering gun pipette tips tip and the bottom of the centrifuge tube, by the centrifuge tube
Peanut leaf tissue mashing, wherein, without visual particulate matter in the peanut leaf tissue after smashing to pieces.
7. a kind of peanut leaf tissue grinder device for being used to extract DNA, it is characterised in that described device includes：
Acquisition module, for taking 0.1 gram ~ 0.4 gram of fresh peanut leaf or taking 0.01 gram ~ 0.04 gram of dry peanut blade,
It is positioned in centrifuge tube, wherein, the moisture content of the fresh peanut leaf is more than 40%, and the moisture content of dry peanut blade is less than
20%, the centrifuge tube is the tubulose sample container of the with closure that volume is 1ml ~ 2ml or gland；
First refrigerating module, for will be equipped with the centrifuge tube of the fresh peanut leaf or the dry peanut blade as
In liquid nitrogen, the temperature of the liquid nitrogen is subzero 196 DEG C；
First processing module, the flame envelope part for the tip of 100ul ~ 300ul liquid-transfering gun pipette tips to be placed in alcolhol burner flame, is treated
After the tip of the liquid-transfering gun pipette tips is melted and turns into ball, by the liquid-transfering gun pipette tips from the flame envelope portion of the alcolhol burner flame
Divide and remove, the tip of the liquid-transfering gun pipette tips is through being cooled into ball, and the radius of the ball is 0.1mm ~ 2mm；
Second refrigerating module, for tip to be positioned over into cooling 3s ~ 10s in liquid nitrogen for the liquid-transfering gun pipette tips of ball；
Second processing module, for the centrifuge tube to be installed for having the fresh flowers leave piece or the dry peanut blade in institute
State when no longer sending sound in liquid nitrogen, the centrifuge tube is taken out from liquid nitrogen be positioned over temperature for subzero 20 DEG C ~ it is subzero 30 DEG C
Centrifugation tube sheet on, and take out from liquid nitrogen the liquid-transfering gun pipette tips, the liquid-transfering gun pipette tips inserted in the centrifuge tube, profit
Quickly the peanut leaf in the centrifuge tube is smashed to pieces with the ball at liquid-transfering gun pipette tips tip, to use the centrifuge tube
The step of carrying out extracting DNA in peanut leaf tissue.
8. device according to claim 7, it is characterised in that the acquisition module specifically for：
Take 0.2 gram of fresh peanut leaf or take 0.02 gram of dry peanut blade, be positioned in centrifuge tube, wherein, it is described from
Heart pipe is conical centrifuge tube, and the bottom inner diameter of the conical centrifuge tube is more than the straight of the sophisticated ball of the liquid-transfering gun pipette tips
Footpath, the specification of the conical centrifuge tube is 1.5ml.
9. the device according to claim 7 or 8, it is characterised in that second refrigerating module specifically for：
By inner chamber of the tip insertion tip of tweezers for the liquid-transfering gun pipette tips of ball, with realize the tweezers are stuck in it is described
Serve as the handle of the liquid-transfering gun pipette tips in the inner chamber of liquid-transfering gun pipette tips；
The mobile tweezers, cooling 3s ~ 4s in liquid nitrogen is positioned over by tip for the liquid-transfering gun pipette tips of ball.
10. the device according to claim 7 or 8, it is characterised in that second refrigerating module specifically for：
Handle is inserted into inner chamber of the tip for the liquid-transfering gun pipette tips of ball, the handle is stuck in the liquid-transfering gun to realize
In the inner chamber of pipette tips, wherein, the inner chamber mutual cooperation of the handle and the liquid-transfering gun pipette tips is realized is stuck in institute by the handle
In the inner chamber for stating liquid-transfering gun pipette tips, the handle is one kind in plastic handle, wood handle or metal handle；
The mobile handle, cooling 3s ~ 4s in liquid nitrogen is positioned over by tip for the liquid-transfering gun pipette tips of ball.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710686110.1A CN107254468A (en) | 2017-08-11 | 2017-08-11 | A kind of peanut leaf tissue grinding methods and device for being used to extract DNA |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710686110.1A CN107254468A (en) | 2017-08-11 | 2017-08-11 | A kind of peanut leaf tissue grinding methods and device for being used to extract DNA |
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| CN107254468A true CN107254468A (en) | 2017-10-17 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201710686110.1A Pending CN107254468A (en) | 2017-08-11 | 2017-08-11 | A kind of peanut leaf tissue grinding methods and device for being used to extract DNA |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107828783A (en) * | 2017-12-13 | 2018-03-23 | 中国农业科学院油料作物研究所 | A kind of peanut leaf DNA highly effective extraction method |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101586102A (en) * | 2009-07-10 | 2009-11-25 | 山东省花生研究所 | Genomic DNA extraction method of peanut leaf blades |
| CN101805730A (en) * | 2009-12-28 | 2010-08-18 | 山东省花生研究所 | Convenient and rapid DNA extracting method of peanut health tissues and diseased tissues |
| CN106434634A (en) * | 2016-10-26 | 2017-02-22 | 遵义医学院 | Method for extracting tissue RNA by tissue grinding |
-
2017
- 2017-08-11 CN CN201710686110.1A patent/CN107254468A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101586102A (en) * | 2009-07-10 | 2009-11-25 | 山东省花生研究所 | Genomic DNA extraction method of peanut leaf blades |
| CN101805730A (en) * | 2009-12-28 | 2010-08-18 | 山东省花生研究所 | Convenient and rapid DNA extracting method of peanut health tissues and diseased tissues |
| CN106434634A (en) * | 2016-10-26 | 2017-02-22 | 遵义医学院 | Method for extracting tissue RNA by tissue grinding |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107828783A (en) * | 2017-12-13 | 2018-03-23 | 中国农业科学院油料作物研究所 | A kind of peanut leaf DNA highly effective extraction method |
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