CN105349272B - A kind of cleaning agent and its preparation method and application method for micropipette needle - Google Patents
A kind of cleaning agent and its preparation method and application method for micropipette needle Download PDFInfo
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- CN105349272B CN105349272B CN201510791398.XA CN201510791398A CN105349272B CN 105349272 B CN105349272 B CN 105349272B CN 201510791398 A CN201510791398 A CN 201510791398A CN 105349272 B CN105349272 B CN 105349272B
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- 239000012459 cleaning agent Substances 0.000 title claims abstract description 83
- 238000000034 method Methods 0.000 title claims abstract description 33
- 238000002360 preparation method Methods 0.000 title claims abstract description 8
- 108020004711 Nucleic Acid Probes Proteins 0.000 claims abstract description 34
- 239000002853 nucleic acid probe Substances 0.000 claims abstract description 34
- 229910021642 ultra pure water Inorganic materials 0.000 claims abstract description 22
- 239000012498 ultrapure water Substances 0.000 claims abstract description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 10
- 239000002904 solvent Substances 0.000 claims abstract description 7
- 239000000523 sample Substances 0.000 claims description 74
- 238000004140 cleaning Methods 0.000 claims description 25
- 239000007788 liquid Substances 0.000 abstract description 47
- 150000007523 nucleic acids Chemical class 0.000 abstract description 12
- 108020004707 nucleic acids Proteins 0.000 abstract description 11
- 102000039446 nucleic acids Human genes 0.000 abstract description 11
- 239000002253 acid Substances 0.000 abstract description 5
- 238000012864 cross contamination Methods 0.000 abstract description 5
- 238000005137 deposition process Methods 0.000 abstract description 3
- 239000004094 surface-active agent Substances 0.000 abstract description 3
- 239000000576 food coloring agent Substances 0.000 abstract description 2
- 238000009396 hybridization Methods 0.000 description 22
- 238000005070 sampling Methods 0.000 description 22
- 238000005406 washing Methods 0.000 description 19
- 239000012224 working solution Substances 0.000 description 16
- 230000000295 complement effect Effects 0.000 description 10
- 239000006210 lotion Substances 0.000 description 9
- 239000011259 mixed solution Substances 0.000 description 9
- 230000000052 comparative effect Effects 0.000 description 8
- 108090000790 Enzymes Proteins 0.000 description 7
- 102000004190 Enzymes Human genes 0.000 description 7
- 238000013019 agitation Methods 0.000 description 6
- 239000000243 solution Substances 0.000 description 5
- 238000001514 detection method Methods 0.000 description 3
- 230000002068 genetic effect Effects 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 238000007605 air drying Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- 235000013399 edible fruits Nutrition 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 238000005457 optimization Methods 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000001394 metastastic effect Effects 0.000 description 1
- 206010061289 metastatic neoplasm Diseases 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000003020 moisturizing effect Effects 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
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Abstract
The invention discloses a kind of cleaning agent for micropipette needle and its preparation method and application method, which includes 0.1 ~ 1%（Mass/volume）Sunset yellow and 0.1 ~ 1%（Volume/volume）TritonX 100, solvent is water.The cleaning agent of the present invention is formulated using food coloring sunset yellow and mild surfactant TritonX 100 by proper proportion, compared to the system liquid of ultra-pure water, the cleaning agent can thoroughly clean up remaining nucleic acid samples, and a variety of different nucleic acid probes is avoided to replace cross contamination caused by deposition process；Compared to strong acid or the cleaning agent of strong basicity, cleaning agent of the invention will not corrode micropipette needle and lead to its lost of life.
Description
Technical field
The present invention relates to micropipette needle technical field more particularly to a kind of cleaning agent for micropipette needle and its match
Method and application method processed.
Background technology
One critical component of non-contacting micro-sampling instrument is piezoelectric type micropipette needle, and the needle is exquisite in workmanship, is easy
Damage, it is expensive.When using such point sample instrument point nucleic acid chip, since nucleic acid samples type often compares more, piezoelectricity
When type micro liquid-transfering needle switches between different samples especially enriched sample, have sample carryover syringe needle inside and outside wall to
The phenomenon that leading to cross contamination between sample.There are cross contamination, nucleic acid chip, that is, unqualified.Solve the problems, such as that this need to be by syringe needle
Cleaning agent, effective syringe needle cleaning agent is less on the market now, and effect is preferable, all has strong acid or strong basicity mostly,
And these strong acid alkaline detergent solutions, which are used for a long time, can cause syringe needle service life to greatly shorten, and virtually increase use cost.
And in such a way that ultra-pure water cleans micropipette needle as system liquid, it tends not to thoroughly clean remaining nucleic acid samples, still deposit
In a degree of cross-contamination phenomena.
Invention content
The present invention provides a kind of cleaning agent for micropipette needle, can thoroughly wash the nucleic acid remained in syringe needle
Sample is again with the moderate characteristic of acid-base property, and the present invention also provides the preparation method of the cleaning agent and application methods.
According to the first aspect of the invention, the present invention provides a kind of cleaning agent for micropipette needle, the cleaning agent packet
The TritonX 100 of the sunset yellow and 0.1~1% (volume/volume) of 0.1~1% (mass/volume) is included, solvent is water.
As the preferred embodiment of the present invention, above-mentioned solvent is ultra-pure water.
As the preferred embodiment of the present invention, above-mentioned cleaning agent includes the TritonX of 0.2~0.8% (volume/volume)
100。
As present invention further optimization scheme, above-mentioned cleaning agent includes 0.3~0.7% (volume/volume)
TritonX 100。
As the still more preferably scheme of the present invention, above-mentioned cleaning agent includes 0.4~0.6% (volume/volume)
TritonX 100。
As the most preferably scheme of the present invention, above-mentioned cleaning agent includes the TritonX 100 of 0.5% (volume/volume).
According to the second aspect of the invention, the present invention provides a kind of preparation method of such as cleaning agent of first aspect, including：
The sunset yellow of formula ratio and TritonX 100 is soluble in water, it is completely dissolved, forms uniform cleaning agent.
According to the third aspect of the invention we, the present invention provides a kind of cleaning agent cleaning micropipette using such as first aspect
The method of needle, including：After above-mentioned micropipette needle pipettes and discharges the sample containing nucleic acid probe, above-mentioned cleaning agent is drawn
To clean above-mentioned micropipette needle.
As the preferred embodiment of the present invention, the time of above-mentioned cleaning is 5-10 seconds.
As present invention further optimization scheme, the time of above-mentioned cleaning is 10 seconds.
The beneficial effects of the invention are as follows：The cleaning agent of the present invention uses food coloring sunset yellow and mild surface-active
Agent TritonX 100 is formulated by proper proportion, compares the system liquid of ultra-pure water, which can thoroughly clean up residual
The nucleic acid samples stayed avoid a variety of different nucleic acid probes from replacing cross contamination caused by deposition process；Compared to strong acid or highly basic
The cleaning agent of property, cleaning agent of the invention will not corrode micropipette needle and lead to its lost of life.
Description of the drawings
Fig. 1 is point master drawing of the nucleic acid probe in comparative example of the present invention and embodiment to chip, wherein first point sample left-hand point,
Then it uses system liquid or cleaning agent to clean micropipette needle, then puts right-hand point, left-hand point and right-hand point respectively illustrate in figure
The color gray scale of sampling liquid；
Fig. 2 is the result figure for cleaning micropipette needle in comparative example of the present invention as system liquid using ultra-pure water, wherein left
Side is that the point sample containing nucleic acid probe is used to work liquid spotting, hybridization and colour developing as a result, right side uses after being cleaned for micropipette needle
Point sample work liquid spotting, hybridization and colour developing result without nucleic acid probe；
Fig. 3 is cleans the result figure of micropipette needle using cleaning agent in the embodiment of the present invention 1, wherein left side contains to use
Point sample work liquid spotting, hybridization and the colour developing of nucleic acid probe are as a result, right side is visited to be used after the cleaning of micropipette needle without nucleic acid
Point sample work liquid spotting, hybridization and the colour developing result of needle.
Specific implementation mode
Below by specific implementation mode combination attached drawing, invention is further described in detail.
The present invention the cleaning agent for micropipette needle, including the sunset yellow of 0.1~1% (mass/volume) with
The TritonX 100 of 0.1~1% (volume/volume), solvent are water.
It should be noted that content of the sunset yellow in cleaning agent is calculated as 0.1~1% according to mass/volume, wherein
The unit of mass/volume is according to the common understanding in this field, such as grams per milliliter (g/mL), such as 0.1~1% (mass/volume)
It can indicate to contain 0.1~1g sunset yellows in 100mL cleaning agents.Similarly, the containing in cleaning agent of TritonX 100
Amount is calculated as 0.1~1% according to volume/volume, the wherein unit of volume/volume according to the common understanding in this field, such as milliliter/
Milliliter (mL/mL), such as 0.1~1% (volume/volume) can indicate to contain 0.1~1mL in 100mL cleaning agents
TritonX 100.Certainly, the above-mentioned explanation to unit is merely exemplary, it is not excluded that using the representation of other units.
TritonX 100 is a kind of mild surfactant, and Chinese is also referred to as " Qula logical -100 ".
Although cleaning agent of the invention can also be however general using other pure water other than ultra-pure water as solvent
Using ultra-pure water commonly used in the art, which has a general concept of this field, such as the ultra-pure water of system liquid,
Therefore its concept is explicit.
It was found that the variation in 0.1~1% (mass/volume) range of content of the sunset yellow in cleaning agent, right
It is influenced without too apparent in cleaning performance；However contents of the TritonX 100 in cleaning agent is in 0.1~1% (volume/body
Product) the interior variation of range, it has a certain impact for cleaning performance.But, contents of the TritonX100 in cleaning agent 0.1~
In 1% (volume/volume) entire scope, preferable cleaning performance can be realized, show the point face that develops the color using INS values as reflection
In the case that color depth is shallow, negative findings are shown, only the specific size of INS values has certain difference, and TritonX 100 is 0.1
In~1% (volume/volume) range, in the case of median, INS values are smaller, illustrate that cleaning performance is better.Therefore,
As the preferred embodiment of the present invention, above-mentioned cleaning agent includes the TritonX 100 of 0.2~0.8% (volume/volume)；As this
The further preferred scheme of invention, above-mentioned cleaning agent include the TritonX 100 of 0.3~0.7% (volume/volume)；As this
The still more preferably scheme of invention, above-mentioned cleaning agent include the TritonX 100 of 0.4~0.6% (volume/volume)；As
The most preferably scheme of the present invention, above-mentioned cleaning agent include the TritonX 100 of 0.5% (volume/volume).
The present invention also provides the preparation methods of above-mentioned cleaning agent, including：By the sunset yellow and TritonX of formula ratio
100 is soluble in water, is completely dissolved, and forms uniform cleaning agent.
Above-mentioned " formula ratio " refers to, prepares the final concentration content of sunset yellow and TritonX 100 in cleaning agent later.
For example, the TritonX 100 of the sunset yellow of 0.1~1% (mass/volume) and 0.1~1% (volume/volume)；0.1~
The TritonX 100 of the sunset yellow of 1% (mass/volume) and 0.2~0.8% (volume/volume)；0.1~1% (quality/
Volume) sunset yellow and 0.3~0.7% (volume/volume) TritonX 100；The day of 0.1~1% (mass/volume)
Fall the TritonX 100 of uranidin and 0.4~0.6% (volume/volume)；The sunset yellow of 0.1~1% (mass/volume)
With the TritonX 100 of 0.5% (volume/volume).
The present invention also provides the methods for using above-mentioned cleaning agent to clean micropipette needle, including：In above-mentioned micropipette
After needle pipettes and discharges the sample containing nucleic acid probe, above-mentioned cleaning agent is drawn to clean above-mentioned micropipette needle.
Method of the method for the cleaning micropipette needle of the present invention relative to existing cleaning micropipette needle, difference are that
With the cleaning agent of the present invention instead of ultra-pure water in the prior art or strong acid or the cleaning agent of strong basicity.As for cleaning procedure
Can be same as the prior art with the time, it is specially required without what.In general, the time of cleaning is 5-10 seconds preferable, more
It is preferred that 10 seconds.
Below by way of comparative example and the embodiment cleaning agent that the present invention will be described in detail and its preparation method and application method, with
And its cleaning performance.
As shown in Figure 1, experimental method is：Two points of point system on chip (or diaphragm), wherein left-hand point are to be visited containing nucleic acid
The point sample working solution of needle, right-hand point are the point sample working solution without nucleic acid probe.Deposition process is：The sample of the complete left-hand point of point sample
Later, system liquid, i.e. ultra-pure water (comparative example) cleaning 10s or cleaning agent using the present invention are respectively adopted to micropipette needle
(embodiment) cleans 10s, then puts the sample of right-hand point processed.
After the completion of point sample, carry out DNA hybridization and colour developing, the hybridization and colour developing can according to this field common practice into
Row.In following embodiment, is hybridized as follows and developed the color.
1DNA hybridizes：
1.1 preparing
1.1.1 prepare reagent
Hybridization neutralizer and washing lotion B are set into 55 DEG C of preheatings.
1.1.2 prepare wet box
It is recommended that production method：With the plastic casing with cover of appropriately sized (being not less than 20cm × 15cm × 10cm), built-in water suction
Paper adds water-soaked, is advisable with no circulating water, 55 DEG C of preheatings.
Pay attention to：Wet box moisturizing must be used in the incubation operation of genetic chip, never wet box is dried！
1.1.3 genetic chip positions
In careful metastatic gene chip to 96 orifice plates, it is placed in chip hole that (normotopia refers to chip level and places, is black by normotopia
Color anchor point is face-up, the lower left corner in the positive device to hole of anchor point；Chip hole refers to the hole position of positioning chip).
Pay attention to：It should avoid area of device contacts chip center when shifting chip and be stained chip！
1.2 prehybridization
100 μ L washing lotions B are added into each chip hole, 55 DEG C incubate 15 minutes；Blot liquid in hole.
1.3 denaturation
30 μ L denaturing liquids are taken to add in PCR product pipe, gentle agitation mixing stands 10 minutes.
1.4 neutralize and hybridize
The PCR product (about 60 μ L) of 100 μ L hybridization neutralizers and denaturation is separately added into each chip hole, gentle agitation is mixed
It is even.
55 DEG C incubate 30 minutes, blot liquid in hole.
1.5 cleaning
150 μ L washing lotion B are added into each chip hole, gentle agitation rinses 3 minutes, blots liquid in hole；It is repeated 2 times.
Pay attention to：Chip temperature in borehole should be made to be not less than 20 DEG C, otherwise influence cleaning performance！
2 colour developings
2.1 prepare reagent
2.1.1 washing lotion A and developing solution B are set into 37 DEG C of preheatings.
2.1.2 enzyme mark working solution is prepared
By 1 requirement of table, enzyme mark stoste is diluted to enzyme mark working solution with washing lotion A.
Pay attention to：It should be included in waste when practical calculating dosage.
Table 1 prepares enzyme mark working solution
| Dosage | Enzyme mark stoste | Washing lotion A |
| 1 part of detection dosage | 1μL | 100μL |
| 24 parts of detection dosages | 24μL | 2.4mL |
2.2 enzyme marks
100 μ L enzyme mark working solutions are taken, are added in the chip hole of cleaning, 37 DEG C incubate 30 minutes；Blot liquid in hole.
2.3 cleaning
150 μ L washing lotion A are added, gentle agitation rinses 3 minutes, blots liquid in hole；It is repeated 1 times.
150 μ L washing lotion C are added, gentle agitation rinses 3 minutes, blots liquid in hole.
Pay attention to：Chip temperature in borehole should be made to be not less than 20 DEG C, otherwise influence cleaning performance！
2.4 colour developing
It is separately added into 50 μ L developing solutions A and 50 μ L developing solution B, gentle agitation mixing, color development at room temperature 5 minutes.
Liquid in hole is blotted, 150 μ L washing lotion C are added, about 1 minute is stood, blots liquid in hole.
2.5 air-drying
45 DEG C air-dry.Pay attention to：PLSCONFM detects after air-drying.
3 detections and analysis
It checks and confirms that genetic chip is normotopia in chip hole, reading system is detected to gene with HPV typing gene chips
Chip is detected and is analyzed.
Comparative example
1,80 μ L sampling liquids are added to (F1 nucleic acid in 5 '-amino labeled nucleic acid probe working solution F1 of isometric 2.4 μM
Probe sequence is：5 '-GCAGACCGAAGTGGATTGCGAGTATTT GA-3 ' come from GeneBank ID：NM_117849), mix
It closes uniform；
2,80 μ L sampling liquids are added in isometric ultra-pure water, are uniformly mixed；
3, two parts of sample RCF9552 in 1 and 2 steps are centrifuged into 5min, the sample centrifuged is loaded respectively to 96 orifice plates
Interior adjacent holes；
4, it elaborates the diaphragm of prescribed level is smooth on the microscope carrier of point sample instrument；
5, point sample instrument parameter is set, setting syringe needle washing procedure to system liquid washs 10s, opens point sample program, point sample
Scheme is as shown in Figure 1；
6, (contain 0.1 μM of 5 '-biotin marks with the nucleic acid solution of the reverse complementary sequence comprising F1 nucleic acid probe sequences
The artificial synthesized nucleic acid fragment of note, sequence are：5’-TCTCTTCCGTTAATTTC
TCAACACATCTTTTCAAATACTCGCAATCCACTTCGGTCTGC-3 ') with chip hybridization and develop the color, record INS values and the moon
Positive findings.
Embodiment 1
1,80 μ L sampling liquids are added in isometric nucleic acid probe working solution F1, are uniformly mixed；
2,80 μ L sampling liquids are added in isometric ultra-pure water, are uniformly mixed；
3, two parts of sample RCF9552 in 1 and 2 are centrifuged into 5min, the sample centrifuged is loaded respectively to 96 orifice plates
Adjacent holes；
4, the mixed solution 100mL works that final concentration is respectively 0.1%TritonX 100 and 0.1% sunset yellow are prepared
For syringe needle cleaning agent；
5, it elaborates the diaphragm of prescribed level is smooth on the microscope carrier of point sample instrument；
6, point sample instrument parameter is set, setting syringe needle washing procedure to syringe needle cleaning agent washs 10s, opens point sample program,
Point sample scheme is as shown in Figure 1；
7, according to above-mentioned hybridizing method, with reverse complementary sequence and chip hybridization and the record INS that develops the color of F1 nucleic acid probes
Value and yin and yang attribute.
Embodiment 2
1,80 μ L sampling liquids are added in isometric nucleic acid probe working solution F1, are uniformly mixed；
2,80 μ L sampling liquids are added in isometric ultra-pure water, are uniformly mixed；
3, two parts of sample RCF9552 in 1 and 2 are centrifuged into 5min, the sample centrifuged is loaded respectively to 96 orifice plates
Adjacent holes；
4, the mixed solution 100mL works that final concentration is respectively 0.1%TritonX 100 and 0.5% sunset yellow are prepared
For syringe needle cleaning agent；
5, it elaborates the diaphragm of prescribed level is smooth on the microscope carrier of point sample instrument；
6, point sample instrument parameter is set, setting syringe needle washing procedure to syringe needle cleaning agent washs 10s, opens point sample program,
Point sample scheme is as shown in Figure 1；
7, according to above-mentioned hybridizing method, with reverse complementary sequence and chip hybridization and the record INS that develops the color of F1 nucleic acid probes
Value and yin and yang attribute.
Embodiment 3
1,80 μ L sampling liquids are added in isometric nucleic acid probe working solution F1, are uniformly mixed；
2,80 μ L sampling liquids are added in isometric ultra-pure water, are uniformly mixed；
3, two parts of sample RCF9552 in 1 and 2 are centrifuged into 5min, the sample centrifuged is loaded respectively to 96 orifice plates
Adjacent holes；
4, the mixed solution 100mL conducts that final concentration is respectively 0.1%TritonX 100 and 1% sunset yellow are prepared
Syringe needle cleaning agent；
5, it elaborates the diaphragm of prescribed level is smooth on the microscope carrier of point sample instrument；
6, point sample instrument parameter is set, setting syringe needle washing procedure to syringe needle cleaning agent washs 10s, opens point sample program,
Point sample scheme is as shown in Figure 1；
7, according to above-mentioned hybridizing method, with reverse complementary sequence and chip hybridization and the record INS that develops the color of F1 nucleic acid probes
Value and yin and yang attribute.
Embodiment 4
1,80 μ L sampling liquids are added in isometric nucleic acid probe working solution F1, are uniformly mixed；
2,80 μ L sampling liquids are added in isometric ultra-pure water, are uniformly mixed；
3, two parts of sample RCF9552 in 1 and 2 are centrifuged into 5min, the sample centrifuged is loaded respectively to 96 orifice plates
Adjacent holes；
4, the mixed solution 100mL works that final concentration is respectively 0.5%TritonX 100 and 0.1% sunset yellow are prepared
For syringe needle cleaning agent；
5, it elaborates the diaphragm of prescribed level is smooth on the microscope carrier of point sample instrument；
6, point sample instrument parameter is set, setting syringe needle washing procedure to syringe needle cleaning agent washs 10s, opens point sample program,
Point sample scheme is as shown in Figure 1；
7, according to above-mentioned hybridizing method, with reverse complementary sequence and chip hybridization and the record INS that develops the color of F1 nucleic acid probes
Value and yin and yang attribute.
Embodiment 5
1,80 μ L sampling liquids are added in isometric nucleic acid probe working solution F1, are uniformly mixed；
2,80 μ L sampling liquids are added in isometric ultra-pure water, are uniformly mixed；
3, two parts of sample RCF9552 in 1 and 2 are centrifuged into 5min, the sample centrifuged is loaded respectively to 96 orifice plates
Adjacent holes；
4, the mixed solution 100mL works that final concentration is respectively 0.5%TritonX 100 and 0.5% sunset yellow are prepared
For syringe needle cleaning agent；
5, it elaborates the diaphragm of prescribed level is smooth on the microscope carrier of point sample instrument；
6, point sample instrument parameter is set, setting syringe needle washing procedure to syringe needle cleaning agent washs 10s, opens point sample program,
Point sample scheme is as shown in Figure 1；
7, according to above-mentioned hybridizing method, with reverse complementary sequence and chip hybridization and the record INS that develops the color of F1 nucleic acid probes
Value and yin and yang attribute.
Embodiment 6
1,80 μ L sampling liquids are added in isometric nucleic acid probe working solution F1, are uniformly mixed；
2,80 μ L sampling liquids are added in isometric ultra-pure water, are uniformly mixed；
3, two parts of sample RCF9552 in 1 and 2 are centrifuged into 5min, the sample centrifuged is loaded respectively to 96 orifice plates
Adjacent holes；
4, the mixed solution 100mL conducts that final concentration is respectively 0.5%TritonX 100 and 1% sunset yellow are prepared
Syringe needle cleaning agent；
5, it elaborates the diaphragm of prescribed level is smooth on the microscope carrier of point sample instrument；
6, point sample instrument parameter is set, setting syringe needle washing procedure to syringe needle cleaning agent washs 10s, opens point sample program,
Point sample scheme is as shown in Figure 1；
7, according to above-mentioned hybridizing method, with reverse complementary sequence and chip hybridization and the record INS that develops the color of F1 nucleic acid probes
Value and yin and yang attribute.
Embodiment 7
1,80 μ L sampling liquids are added in isometric nucleic acid probe working solution F1, are uniformly mixed；
2,80 μ L sampling liquids are added in isometric ultra-pure water, are uniformly mixed；
3, two parts of sample RCF9552 in 1 and 2 are centrifuged into 5min, the sample centrifuged is loaded respectively to 96 orifice plates
Adjacent holes；
4, the mixed solution 100mL conducts that final concentration is respectively 1%TritonX 100 and 0.1% sunset yellow are prepared
Syringe needle cleaning agent；
5, it elaborates the diaphragm of prescribed level is smooth on the microscope carrier of point sample instrument；
6, point sample instrument parameter is set, setting syringe needle washing procedure to syringe needle cleaning agent washs 10s, opens point sample program,
Point sample scheme is as shown in Figure 1；
7, according to above-mentioned hybridizing method, with reverse complementary sequence and chip hybridization and the record INS that develops the color of F1 nucleic acid probes
Value and yin and yang attribute.
Embodiment 8
1,80 μ L sampling liquids are added in isometric nucleic acid probe working solution F1, are uniformly mixed；
2,80 μ L sampling liquids are added in isometric ultra-pure water, are uniformly mixed；
3, two parts of sample RCF9552 in 1 and 2 are centrifuged into 5min, the sample centrifuged is loaded respectively to 96 orifice plates
Adjacent holes；
4, the mixed solution 100mL conducts that final concentration is respectively 1%TritonX 100 and 0.5% sunset yellow are prepared
Syringe needle cleaning agent；
5, it elaborates the diaphragm of prescribed level is smooth on the microscope carrier of point sample instrument；
6, point sample instrument parameter is set, setting syringe needle washing procedure to syringe needle cleaning agent washs 10s, opens point sample program,
Point sample scheme is as shown in Figure 1；
7, according to above-mentioned hybridizing method, with reverse complementary sequence and chip hybridization and the record INS that develops the color of F1 nucleic acid probes
Value and yin and yang attribute.
Embodiment 9
1,80 μ L sampling liquids are added in isometric nucleic acid probe working solution F1, are uniformly mixed；
2,80 μ L sampling liquids are added in isometric ultra-pure water, are uniformly mixed；
3, two parts of sample RCF9552 in 1 and 2 are centrifuged into 5min, the sample centrifuged is loaded respectively to 96 orifice plates
Adjacent holes；
4, it is respectively the mixed solution 100mL of 1%TritonX 100 and 1% sunset yellow as needle to prepare final concentration
Head cleaning agent；
5, it elaborates the diaphragm of prescribed level is smooth on the microscope carrier of point sample instrument；
6, point sample instrument parameter is set, setting syringe needle washing procedure to syringe needle cleaning agent washs 10s, opens point sample program,
Point sample scheme is as shown in Figure 1；
7, according to above-mentioned hybridizing method, with reverse complementary sequence and chip hybridization and the record INS that develops the color of F1 nucleic acid probes
Value and yin and yang attribute.
The colour developing result of comparative example and embodiment 1 difference is as shown in Figures 2 and 3.Fig. 2 is using ultra-pure water as system liquid
The result figure of micropipette needle is cleaned, wherein left side is using point sample work liquid spotting, hybridization and colour developing knot containing nucleic acid probe
Fruit, right side are after micropipette needle cleans using point sample work liquid spotting, hybridization and colour developing result without nucleic acid probe.Fig. 2
The results show that the point on right side has obvious colour developing gray scale, show that cleaning performance is bad.Fig. 3 is to be cleaned using cleaning agent
The result figure of micropipette needle, wherein left side is to work liquid spotting, hybridization and colour developing as a result, the right side using the point sample containing nucleic acid probe
Side is after micropipette needle cleans using point sample work liquid spotting, hybridization and colour developing result without nucleic acid probe.The result of Fig. 3
The point of display, right side does not observe apparent colour developing gray scale, shows that cleaning performance is good.
Table 2 shows the INS values and yin and yang attribute result that the above comparative example and embodiment obtain.
The INS values and yin and yang attribute that 2 comparative example of table and embodiment obtain
Note：An INS value sizes reflection colour developing point shade, when Zhi≤11 INS are judged as positive findings,<11 be negative knot
Fruit.
The above content is combining, specific embodiment is made for the present invention to be further described, and it cannot be said that this hair
Bright specific implementation is confined to these explanations.For those of ordinary skill in the art to which the present invention belongs, it is not taking off
Under the premise of from present inventive concept, a number of simple deductions or replacements can also be made, all shall be regarded as belonging to the protection of the present invention
Range.
Claims (10)
1. a kind of cleaning agent for micropipette needle, which is characterized in that the cleaning agent includes 0.1 ~ 1%（Mass/volume）
Sunset yellow and 0.1 ~ 1%（Volume/volume）TritonX 100, solvent is water.
2. cleaning agent according to claim 1, which is characterized in that the solvent is ultra-pure water.
3. cleaning agent according to claim 1 or 2, which is characterized in that the cleaning agent includes 0.2 ~ 0.8%（Volume/body
Product）TritonX 100.
4. cleaning agent according to claim 1 or 2, which is characterized in that the cleaning agent includes 0.3 ~ 0.7%（Volume/body
Product）TritonX 100.
5. cleaning agent according to claim 1 or 2, which is characterized in that the cleaning agent includes 0.4 ~ 0.6%（Volume/body
Product）TritonX 100.
6. cleaning agent according to claim 1 or 2, which is characterized in that the cleaning agent includes 0.5%（Volume/volume）
TritonX 100.
7. a kind of preparation method of cleaning agent as claimed in any one of claims 1 to 6, which is characterized in that by the day of formula ratio
It falls uranidin and TritonX 100 is soluble in water, be completely dissolved, form uniform cleaning agent.
8. a kind of method for cleaning micropipette needle using cleaning agent as claimed in any one of claims 1 to 6, which is characterized in that
After the micropipette needle pipettes and discharges the sample containing nucleic acid probe, it is described micro to clean to draw the cleaning agent
Liquid-transfering needle.
9. according to the method described in claim 8, it is characterized in that, the time of the cleaning is 5-10 seconds.
10. according to the method described in claim 9, it is characterized in that, the time of the cleaning is 10 seconds.
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| CN112048401B (en) * | 2020-08-28 | 2022-05-17 | 上海符贝基因科技有限公司 | Micro-fluidic chip cleaning agent and method thereof |
| CN115044417A (en) * | 2022-05-11 | 2022-09-13 | 苏州莱博睿思生物科技有限公司 | Cleaning fluid, preparation method and application thereof |
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| CN101939427A (en) * | 2008-01-18 | 2011-01-05 | 国立大学法人富山大学 | Reaction device, reaction method and method of synthesizing cDNA |
| CN103930546A (en) * | 2011-09-26 | 2014-07-16 | 凯杰有限公司 | Rapid method for isolating extracellular nucleic acids |
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| CN1566304A (en) * | 2003-06-13 | 2005-01-19 | 李小平 | Full automatic biochemical analyzer cleaning agent |
| US20060223070A1 (en) * | 2005-04-01 | 2006-10-05 | Wisniewski Michele E | Methods, compositions, and kits for detecting nucleic acids in a single vessel |
| JP4361052B2 (en) * | 2005-12-26 | 2009-11-11 | ライオン株式会社 | Liquid detergent composition |
| AU2010351576B2 (en) * | 2010-04-23 | 2014-08-14 | FinaBioSolutions, LLC | Simple method for simultaneous removal of multiple impurities from culture supernatants to ultralow levels |
| US20130005591A1 (en) * | 2011-01-13 | 2013-01-03 | Halgen Corporation | Method for parallel amplification of nucleic acids |
| CN104673511A (en) * | 2013-11-30 | 2015-06-03 | 青岛科润塑料机械有限公司 | Novel detergent |
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| CN103930546A (en) * | 2011-09-26 | 2014-07-16 | 凯杰有限公司 | Rapid method for isolating extracellular nucleic acids |
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