CN110343738A - The Method and kit for of costal cartilage DNA is acquired and is stored at room temperature based on FTA card - Google Patents
The Method and kit for of costal cartilage DNA is acquired and is stored at room temperature based on FTA card Download PDFInfo
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- CN110343738A CN110343738A CN201910523371.0A CN201910523371A CN110343738A CN 110343738 A CN110343738 A CN 110343738A CN 201910523371 A CN201910523371 A CN 201910523371A CN 110343738 A CN110343738 A CN 110343738A
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- QTCANKDTWWSCMR-UHFFFAOYSA-N costic aldehyde Natural products C1CCC(=C)C2CC(C(=C)C=O)CCC21C QTCANKDTWWSCMR-UHFFFAOYSA-N 0.000 title claims abstract description 86
- ISTFUJWTQAMRGA-UHFFFAOYSA-N iso-beta-costal Natural products C1C(C(=C)C=O)CCC2(C)CCCC(C)=C21 ISTFUJWTQAMRGA-UHFFFAOYSA-N 0.000 title claims abstract description 86
- 238000000034 method Methods 0.000 title claims abstract description 33
- 210000001519 tissue Anatomy 0.000 claims abstract description 19
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 8
- 238000005520 cutting process Methods 0.000 claims abstract description 4
- 210000000003 hoof Anatomy 0.000 claims abstract description 4
- 244000137852 Petrea volubilis Species 0.000 claims description 17
- 238000003801 milling Methods 0.000 claims description 7
- 239000004576 sand Substances 0.000 claims description 7
- IAYPIBMASNFSPL-UHFFFAOYSA-N Ethylene oxide Chemical compound C1CO1 IAYPIBMASNFSPL-UHFFFAOYSA-N 0.000 claims description 4
- 239000004568 cement Substances 0.000 claims description 4
- 239000003153 chemical reaction reagent Substances 0.000 claims description 4
- 230000020477 pH reduction Effects 0.000 claims description 3
- 238000004321 preservation Methods 0.000 abstract description 13
- 239000000463 material Substances 0.000 abstract description 7
- 238000000605 extraction Methods 0.000 abstract description 6
- DNJIEGIFACGWOD-UHFFFAOYSA-N ethanethiol Chemical compound CCS DNJIEGIFACGWOD-UHFFFAOYSA-N 0.000 abstract description 4
- 108020004414 DNA Proteins 0.000 description 32
- 238000007689 inspection Methods 0.000 description 11
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 7
- 238000001514 detection method Methods 0.000 description 7
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Abstract
The present invention is based on methods FTA card acquisition and be stored at room temperature costal cartilage DNA, which is characterized in that its step includes: S1. acquisition corpse costal cartilage 1-2cm, and knot hoof tissue and the corrupt tissue around the corpse costal cartilage are rejected with the scalpel cleaned by pure water；S2. taking in step S1 that treated, corpse costal cartilage is ground, and is instilled pure water and mixed, and tissue homogenate is prepared into；S3. it takes the tissue prepared in suitable step S2 homogenate to be applied within the scope of the annulus indicated on FTA card with sponge label are viscous, prepares FTA sample card, and dried in the shade；S4. the FTA sample prepared in step S3 is snapped fits into cutting ferrule and hermetic bag, the hermetic bag is vacuumized, and preservation is sealed.The method that the present invention can be realized extraction DNA is easy to operate easily stored with material, so that having saved extraction time, also avoids corrupt stench situation, reduces the health risk of staff.
Description
Technical field
The present invention relates to a kind of technical fields based on FTA card acquisition DNA, are based on FTA card more specifically to one kind
Acquire and be stored at room temperature the Method and kit for of costal cartilage DNA.
Background technique
In the daily inspection case of forensic, when the tissues such as the painstaking effort of decomposed body, muscle can not detect short STR result,
It extracts its costal cartilage progress STR parting and frequently results in satisfied effect, it is one that corpse costal cartilage tissue, which is not easy corruption compared with soft tissue,
Kind common biological material, in the cases such as disaster accident, the dead group's wound of group, not well-known corpse frequently as preferred biological material into
Row extracts.
The research and application of Forensic DNA typing are initially since length polymorphism, and in early stage, Forensic DNA typing is answered
The site moonlet VNTR is detected with restriction fragment length polymorphism (RFLP) analytical technology, constructs DNA fingerprinting, the nineties
After mid-term, it is gradually transitions the polymorphism using the round pcr analysis seat microsatellite STR, i.e. STR typing method.STR is current
Conventional use of length polymorphism genetic marker in forensic identification, allele fragment length is mostly in 400bp hereinafter, adopt
High with round pcr amplification success rate, positive rate and detection sensitivity are about 10 times higher than moonlet VNTR, are particularly suitable for dropping
The Classification Identification of solution, outmoded and corrupt sample.
Corpse costal cartilage group is extracted at the scene generally by staff to the STR parting detection of costal cartilage tissue at present
After knitting, refrigeration is sent to laboratory, is then extracted by the appraiser in laboratory using Chelex100 method and obtains DNA, as PCR
The template of test.However, this method extracts DNA complex steps, time-consuming, is frequently accompanied by stench, disinfect pathogen on corpse costal cartilage
Proliferation has easily led to live extraction personnel health and has been on the hazard, and inspection needs freezen protective, and accumulation occupies a large amount of refrigerations throughout the year
Reefer space, once power-off or there is refrigerating equipment failure, costal cartilage can accelerate corruption to even result in sample to loss.
FTA (Flinders Technology Associates) card is Whatman house journal, Britain product, is room temperature
Lower collection, transport and stored DNA provide safe, safe and reliable method.Some researches show that FTA card can be used as a kind of logical
With, DNA profiling preparation method quickly, easy, containing special chemical substance in FTA card, when sample is adsorbed on card, carefully
After birth and organelle are cleaved, protein denaturation, and the nucleic acid released is trapped on FTA card fiber, thus from nuclease,
Oxidant and ultraviolet injury.FTA card can rapidly be such that organism (including the various pathogen in blood) inactivate, and press down
The growth of bacterium processed and other microorganisms, genomic DNA can store under room temperature 17 years or more on FTA card, without dropping
Low amplification efficiency.In addition, the FTA gene preservation system of u s company's research and development, which is stored at room temperature aspect in gene, also good effect
Fruit can save gene samples for a long time, in long distance transportation gene samples and regardless of conditions pair such as environment temperatures
The influence of gene samples.
The present invention establishes a kind of Method and kit for for extracting based on FTA card and acquiring and be stored at room temperature costal cartilage DNA, so that will
The method that costal cartilage is prepared into costal cartilage FTA card is stored in costal cartilage on FTA card, close by direct expansion method and vacuum Material evidence bag
Envelope realizes that costal cartilage rapid DNA is examined and room temperature long-term preservation.The present invention can substitute the cumbersome of traditional costal cartilage DNA extraction
Step can solve costal cartilage tissue and be difficult to the problem saved at normal temperature.
Summary of the invention
The present invention overcomes the deficiencies in the prior art, provide one kind and are acquired and be stored at room temperature costal cartilage DNA based on FTA card
Method and kit for, can be realized extract costal cartilage DNA method is easy to operate and material is easily stored so that having saved extraction
Time avoids corrupt stench situation, reduces the health risk of staff.
To achieve the goals above, present invention employs following technical proposals:
The method for being acquired based on FTA card and being stored at room temperature costal cartilage DNA, step include:
S1. corpse costal cartilage 1-2cm is acquired, is rejected around the corpse costal cartilage with the scalpel cleaned by pure water
Knot hoof tissue and corrupt tissue；
S2. taking in step S1 that treated, corpse costal cartilage is ground, and is instilled pure water and mixed, and is prepared into
Tissue homogenate；
S3. the tissue prepared in suitable step S2 homogenate is taken to be applied within the scope of the annulus indicated on FTA card with sponge label are viscous,
FTA sample card is prepared, and is dried in the shade；
S4. the FTA sample prepared in step S3 being snapped fits into cutting ferrule and hermetic bag, the hermetic bag is vacuumized,
It is sealed preservation.
The present invention also provides the tools that corpse costal cartilage in a kind of step S2 in claim 1 is ground, it is wrapped
It includes including plastic plate and sticks in the sand paper in the plastic cement plate end face, the plastic plate and sand paper carry out at nucleination
Reason.
As a further explanation of the present invention, the plastic cement plate end face includes first treatment region and sand face milling zone, the sand
Paper sticks on the milling zone of the sand face.
As a further explanation of the present invention, the roughness of the sand paper is 400 mesh.
As a further explanation of the present invention, the plastic plate and sand paper pass through ethylene oxide reagent and carry out nucleic acid
Change processing.
As a further explanation of the present invention, the plastic plate and sand paper pass through ethylene oxide reagent and carry out nucleic acid
Change processing.
Compared with prior art, beneficial effects of the present invention:
The tedious steps that the present invention is extracted instead of DNA, operation of the present invention is simple, when saving the work of Testing and appraisal
Between, workload is reduced, is improved work efficiency；The homogenate of costal cartilage tissue is acquired by FTA card, corpse is avoided and rots to dislike
It is smelly, meanwhile, the chance that harmful sick body is propagated is also reduced, staff's health risk is reduced；It is soft with FTA card acquisition corpse rib
Bone tissue homogenate, can room temperature storage, taking back laboratory can be directly used for subsequent PCR detection, eliminate the extraction work of DNA, material
Material is also easy to store, and store method is simple, meanwhile, but also convenient transport；Tool by preparing the homogenate of costal cartilage tissue will
Costal cartilage is ground into tissue homogenate, and is adsorbed on FTA card, low in cost, easy to operate, and makes its Sample preservation more
Securely and reliably, clean tasteless, transport inspection easy to carry.
Detailed description of the invention
Fig. 1 is that costal cartilage FTA card of the invention directly expands obtained STR parting figure；
Fig. 2 is the STR parting figure of control group costal cartilage of the present invention；
Fig. 3 is that the present invention 2 years costal cartilage FTA cards of preservation directly expand obtained STR parting figure；
Fig. 4 is the structural schematic diagram for the tool that the present invention prepares the homogenate of costal cartilage tissue.
Specific embodiment
Further describe with reference to the accompanying drawings and detailed description to the present invention: following embodiment is for illustrating this hair
It is bright, but do not have to limit the scope of the invention.Unless otherwise specified, embodiment is according to " forensic DNA profiling laboratory inspection specification
(GA/T383-2014) " experiment condition as defined in, or the condition according to manufacturer's specification suggestion.As pcr amplification reaction makes
Reaction system is operated by used amplification tool (the identification of Forensic DNA kit) specification.
FTA card used in the following embodiment is purchased from Britain Whatman company, Identifiler Plus amplification kit
Purchased from Thermo Fisher company, the U.S..
In the present embodiment, test sample is 158 parts of costal cartilage sample of daily inspection case accumulation, and wherein fresh cadaver rib is soft
130 parts of bone, 17 parts of slight decomposed body costal cartilage, 11 parts of height decomposed body costal cartilage.
The method extracted based on FTA card and be stored at room temperature costal cartilage DNA, step include:
S1. corpse costal cartilage 1-2cm is acquired, is rejected around the corpse costal cartilage with the scalpel cleaned by pure water
Knot hoof tissue and corrupt tissue；
S2. taking in step S1 that treated, corpse costal cartilage is ground, and is instilled pure water and mixed, and is prepared into
Tissue homogenate；
S3. the tissue prepared in suitable step S2 homogenate is taken to be applied within the scope of the annulus indicated on FTA card with sponge label are viscous,
The homogenate of costal cartilage tissue is just adsorbed on FTA card, prepares FTA sample card, in this present embodiment, is prepared according to different items
3 class FTA samples of classification production under part (fresh cadaver costal cartilage, slight decomposed body costal cartilage and height decomposed body costal cartilage)
Card sample, and dried in the shade；
S4. the FTA sample prepared in step S3 being snapped fits into cutting ferrule and hermetic bag, the hermetic bag is vacuumized,
It is sealed preservation.
After drying, can room temperature preservation, Sample preservation is more safe and reliable, clean tasteless, transport inspection easy to carry.
Sample is sent to laboratory, eliminates the process for extracting DNA, it is also test sensitivity section that laboratory worker, which avoids stench,
About time.
In the present embodiment, 3 class FTA sample cards are carried out amplification reaction, amplified reaction uses IdentifilerPlus
Amplification kit amplification, 25 μ L of amplified reaction total volume, thermal cycle conditions are set according to kit recommended parameter, then, are expanded
Volume increase object carries out electrophoresis detection through ABI-3500XL type genetic analyzer and analysis, electrophoresis provide to carry out to specifications, with
GeneMapper ID-X software makees STR phenotypic analysis, obtains such as Fig. 1-3.
By test and the discovery of STR parting, 158 parts of costal cartilage tissues in this test are mentioned by Chelex-100 method
It takes, has obtained complete STR parting, after preparing FTA card respectively, directly expand, there are 147 parts of costal cartilage FTA cards to obtain complete
Whole STR parting, genotyping result is good, and STR recall rate is 93.0% (147/158)；It is certain case with reference to Fig. 2
A201609097008 sample directly expands obtained STR map using costal cartilage FTA card, is with reference to Fig. 3
The STR map that A201609097008 sample is expanded after being extracted using Chelex-100 method, two kinds of sides of same sample
Method detection and genotyping result is consistent, has no allelic loss phenomenon, in addition, 11 parts of costal cartilage FTA of effective STR parting are not detected
Card belongs to height decomposed body costal cartilage sample.
In another test, (147 parts) of costal cartilage FTA card effectively detected are sealed with vacuum Material evidence bag, number, room temperature
It is lower to save 2 years, it is punched on FTA card using 2mm punch, takes 1 for PCR direct expansion, shown by experimental study
After the upper costal cartilage FTA card (147 parts) effectively detected saves 2 years at normal temperature, carry out punching direct augmentation detection, STR
Recall rate is 100%, and parting is good, has no allelic loss phenomenon, is that A201609097008 sample is adopted with reference to Fig. 4
The STR map directly expanded after being saved 2 years with costal cartilage FTA card, by comparison diagram 2 and Fig. 4, rigid manufactured costal cartilage
FTA card, which directly expands, relatively to be saved 2 years costal cartilage FTA cards and directly expands, gained map in PCR product peak height fluctuation range, together
It is more preferable in the indexs such as peak equalization between heterozygote on one locus.
Costal cartilage is the common sample in postmortem examination, and it is to not well-known corpse that DNA is extracted from costal cartilage and examines STR parting
The important means of body progress identification of corpse.This method is stored at room temperature costal cartilage tissue using FTA card, relative to other preservation sides
Formula, this method may be implemented to be stored at room temperature.Currently, costal cartilage is frequently with freezen protective, this method preservation sample difficulty is big, fortune
Such as saved during defeated it is improper easily cause sample corrupt, lead to DNA degradation.If needed being saved after costal cartilage shave
Costal cartilage thin slice is dried, this method is more demanding to sample Conservation environment, is not suitable for that southern humidity is big, temperature is high
Climatic environment.Relative to traditional inspection mode, the present embodiment can room temperature inspection, safe ready, especially costal cartilage FTA card
Preparation is not limited by time, region, can be completed in regions such as criminal-scene, autopsy room, laboratories, inspection
The propagation of harmful pathogen is avoided in journey.Meanwhile now surveying personnel and being not necessarily to the inspection of costal cartilage tissue to the laboratory DNA, thus
The cleaning health that ensure that the laboratory DNA, it is possible to prevente effectively from sample mutually pollutes, it is possible to reduce the harm to reviewer.
In addition, DNA reviewer, after receiving costal cartilage FTA card, punching directly expands, cumbersome digestion is eliminated
Extraction time is greatly saved in process, during the experiment, can by the size in punch aperture and the number of the punching scraps of paper come
Control the template DNA amount of amplification.Meanwhile costal cartilage tissue saved using FTA card not influence DNA secondarily purified, it is more for impurity
Sample, can be rapidly purified by WATER-WASHING METHOD, also can be used other methods such as silica bead method to costal cartilage DNA carry out it is secondarily purified,
To obtain higher-quality template DNA [12].100% can be reached by being stored at room temperature 2 years costal cartilage FTA card recall rates, and be divided
Type is good, has no allelic loss phenomenon, it was confirmed that FTA card saves the validity of costal cartilage DNA, has met daily material evidence and has protected
Deposit the demand of inspection.Certainly, can there are degradation, comparison diagram 2 and Fig. 4 during preservation due to DNA, just manufactured costal cartilage
FTA card, which directly expands, relatively to be saved 2 years costal cartilage FTA cards and directly expands, gained map in PCR product peak height fluctuation range, together
It is more preferable in the indexs such as peak equalization between heterozygote on one locus.158 parts of costal cartilage tissues in the present embodiment pass through routine
Method (Chelex-100 method) is extracted, and has been obtained complete STR parting, has been transferred completely on FTA card, is directly expanded,
There are 147 parts of costal cartilage FTA cards to obtain complete STR parting, there are 11 parts of FTA cards that effective STR parting is not detected, this 11 parts
Belong to height decomposed body costal cartilage.
Costal cartilage tissue is saved using FTA card, the limitation of cryo-conservation can be overcome, is mentioned for long distance transportation under room temperature
Condition has been supplied, the validity of preservation and the safety of transport are improved.So far, costal cartilage tissue can save on FTA card
At least 2 years.
With reference to Fig. 4, in an alternative embodiment of the invention, additionally provides and made in a kind of step S1 in claim 1
The tool that standby costal cartilage tissue is homogenized, also referred to as disposable tissue homogenate are frustrated, it includes including plastic plate 1 and sticking in the modeling
Sand paper 2 on 1 end face of offset plate, the plastic plate and sand paper 2 carry out stoning acidification.
In the present embodiment, described preferably, 1 end face of the plastic plate includes first treatment region 3 and sand face milling zone 4
Sand paper 2 sticks on sand face milling zone 4, in addition, the just area ratio most preferably 1:1 for the treatment of region 3 and sand face milling zone 4.
In the present embodiment, preferably, the roughness of the sand paper 2 is 400 mesh, the sand paper 2 of this 400 mesh roughness
Go grinding corpse costal cartilage better effect.
In the present embodiment, preferably, the plastic plate and sand paper 2 carries out nucleic acid by ethylene oxide reagent
Change processing prevents pollution corpse costal cartilage, increases detection precision.
Claims (5)
1. being acquired based on FTA card and the method that is stored at room temperature costal cartilage DNA, which is characterized in that its step includes:
S1. corpse costal cartilage 1-2cm is acquired, the knot around the corpse costal cartilage is rejected with the scalpel cleaned by pure water
Hoof tissue and corrupt tissue；
S2. taking in step S1 that treated, corpse costal cartilage is ground, and is instilled pure water and mixed, and tissue is prepared into
Homogenate；
S3. it takes the tissue prepared in suitable step S2 homogenate to be applied within the scope of the annulus indicated on FTA card with sponge label are viscous, prepares
FTA sample card, and dried in the shade；
S4. the FTA sample prepared in step S3 is snapped fits into cutting ferrule and hermetic bag, the hermetic bag is vacuumized, and is carried out
It is sealed.
2. the tool that the corpse costal cartilage in a kind of step S2 in claim 1 is ground, which is characterized in that it includes packet
It includes plastic plate and sticks in the sand paper in the plastic cement plate end face, the plastic plate and sand paper carry out stoning acidification.
3. the tool according to claim 2 for preparing the homogenate of costal cartilage tissue, which is characterized in that the plastic cement plate end face packet
Just treatment region and sand face milling zone, the sand paper is included to stick on the milling zone of the sand face.
4. the tool according to claim 2 or 3 for preparing the homogenate of costal cartilage tissue, which is characterized in that the sand paper
Roughness is 400 mesh.
5. the tool according to claim 2 for preparing the homogenate of costal cartilage tissue, which is characterized in that the plastic plate and sand
Paper passes through ethylene oxide reagent and carries out stoning acidification.
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| CN113005173A (en) * | 2019-12-19 | 2021-06-22 | 武汉艾米森生命科技有限公司 | Feces exfoliated cell nucleic acid preservation reagent and preparation method and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2002238558A (en) * | 2001-02-13 | 2002-08-27 | Dna Bank:Kk | Method for preserving dna and method for obtaining therapeutic information by making use of the thus preserved dna |
| CN101305708A (en) * | 2008-07-09 | 2008-11-19 | 长春市博坤生物科技有限公司 | Long term storage process of biological sample |
| CN103698280A (en) * | 2013-12-10 | 2014-04-02 | 汕头大学医学院 | Method for fixing and extracting biological tissue by using immobile liquid to deduce death time of biological tissues |
| CN104164496A (en) * | 2014-07-24 | 2014-11-26 | 中国农业科学院棉花研究所 | Method for plant PCR detection based on preservation of DNA in FTA card |
| US20160017315A1 (en) * | 2013-03-15 | 2016-01-21 | Ge Healthcare Uk Limited | Methods for one step nucleic acid amplification of non-eluted samples |
| CN108118049A (en) * | 2016-11-28 | 2018-06-05 | 上海怡泽生物科技有限公司 | The method of one rapid extraction RNA from animal tissue |
-
2019
- 2019-06-17 CN CN201910523371.0A patent/CN110343738A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2002238558A (en) * | 2001-02-13 | 2002-08-27 | Dna Bank:Kk | Method for preserving dna and method for obtaining therapeutic information by making use of the thus preserved dna |
| CN101305708A (en) * | 2008-07-09 | 2008-11-19 | 长春市博坤生物科技有限公司 | Long term storage process of biological sample |
| US20160017315A1 (en) * | 2013-03-15 | 2016-01-21 | Ge Healthcare Uk Limited | Methods for one step nucleic acid amplification of non-eluted samples |
| CN103698280A (en) * | 2013-12-10 | 2014-04-02 | 汕头大学医学院 | Method for fixing and extracting biological tissue by using immobile liquid to deduce death time of biological tissues |
| CN104164496A (en) * | 2014-07-24 | 2014-11-26 | 中国农业科学院棉花研究所 | Method for plant PCR detection based on preservation of DNA in FTA card |
| CN108118049A (en) * | 2016-11-28 | 2018-06-05 | 上海怡泽生物科技有限公司 | The method of one rapid extraction RNA from animal tissue |
Non-Patent Citations (5)
| Title |
|---|
| 平安清远: "软骨组织采集与保存套装", 《HTTPS://MP.WEIXIN.QQ.COM/S?SRC=3&TIMESTAMP=1667272193&VER=1&SIGNATURE=Y3QJSP3LAES5SRRENE*N6O*2WM6XNQPCYK5UMIRQPYEUEON*XHOCGH3988CC6IGS49GQ9ZOOZPEVNESF7EWAAACMSGTH5PUBBSMV3X-QC*ZGEPRUN6CVFPBC*5CHAFOW5YUGVG*WWRUHYWSAHRIKE0WDMZLXJWMIYTKXS1JFZ8W=》 * |
| 李安等: "多种载体人体生物检材直扩方法在DNA数据库建库中的实验研究", 《昆明医学院学报》 * |
| 李林等: "直接扩增法在血痕、肋软骨和唾液检材中的应用", 《中国法医学杂志》 * |
| 胡萌等: "19个常染色体STR复合扩增体系在实际案件中的应用探析", 《铁道警察学院学报》 * |
| 邹凯南等: "Expressmarker 22 STR荧光检测试剂盒的法医学应用", 《法医学杂志》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113005173A (en) * | 2019-12-19 | 2021-06-22 | 武汉艾米森生命科技有限公司 | Feces exfoliated cell nucleic acid preservation reagent and preparation method and application thereof |
| CN113005173B (en) * | 2019-12-19 | 2023-10-27 | 武汉艾米森生命科技有限公司 | Fecal exfoliated cell nucleic acid preservation reagent and preparation method and application thereof |
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