CN1112124A - Process for extracting deoxyribonucleic acid (DNA) - Google Patents
Process for extracting deoxyribonucleic acid (DNA) Download PDFInfo
- Publication number
- CN1112124A CN1112124A CN 94110993 CN94110993A CN1112124A CN 1112124 A CN1112124 A CN 1112124A CN 94110993 CN94110993 CN 94110993 CN 94110993 A CN94110993 A CN 94110993A CN 1112124 A CN1112124 A CN 1112124A
- Authority
- CN
- China
- Prior art keywords
- dna
- precipitation
- nucleic acid
- extraction process
- spermary
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 102000053602 DNA Human genes 0.000 title abstract description 19
- 108020004414 DNA Proteins 0.000 title abstract description 19
- 238000000034 method Methods 0.000 title abstract description 8
- 238000000605 extraction Methods 0.000 claims abstract description 13
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims abstract description 12
- 241000251468 Actinopterygii Species 0.000 claims abstract description 11
- 239000000047 product Substances 0.000 claims abstract description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 8
- 238000003756 stirring Methods 0.000 claims abstract description 8
- 239000001509 sodium citrate Substances 0.000 claims abstract description 7
- 235000017557 sodium bicarbonate Nutrition 0.000 claims abstract description 6
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims abstract description 6
- 238000001556 precipitation Methods 0.000 claims description 10
- 210000001541 thymus gland Anatomy 0.000 claims description 9
- 102000039446 nucleic acids Human genes 0.000 claims description 8
- 108020004707 nucleic acids Proteins 0.000 claims description 8
- 150000007523 nucleic acids Chemical class 0.000 claims description 8
- 239000000243 solution Substances 0.000 claims description 6
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 claims description 6
- 229940038773 trisodium citrate Drugs 0.000 claims description 5
- 235000013305 food Nutrition 0.000 claims description 4
- 239000002994 raw material Substances 0.000 claims description 4
- 239000000835 fiber Substances 0.000 claims description 3
- 230000007062 hydrolysis Effects 0.000 claims description 3
- 238000006460 hydrolysis reaction Methods 0.000 claims description 3
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 claims description 2
- 238000001035 drying Methods 0.000 claims description 2
- 239000012535 impurity Substances 0.000 claims description 2
- 239000011259 mixed solution Substances 0.000 claims description 2
- 239000006228 supernatant Substances 0.000 claims description 2
- 102000004190 Enzymes Human genes 0.000 claims 1
- 108090000790 Enzymes Proteins 0.000 claims 1
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 claims 1
- 239000002253 acid Substances 0.000 claims 1
- 230000008030 elimination Effects 0.000 claims 1
- 238000003379 elimination reaction Methods 0.000 claims 1
- 238000009413 insulation Methods 0.000 claims 1
- 210000000496 pancreas Anatomy 0.000 claims 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 abstract description 4
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 4
- 238000004519 manufacturing process Methods 0.000 abstract description 4
- 239000000463 material Substances 0.000 abstract description 4
- 238000003912 environmental pollution Methods 0.000 abstract description 3
- 150000003839 salts Chemical class 0.000 abstract description 3
- 108010019160 Pancreatin Proteins 0.000 abstract description 2
- 229940055695 pancreatin Drugs 0.000 abstract description 2
- 239000002244 precipitate Substances 0.000 abstract description 2
- 239000011780 sodium chloride Substances 0.000 abstract description 2
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 abstract description 2
- 231100000331 toxic Toxicity 0.000 abstract description 2
- 230000002588 toxic effect Effects 0.000 abstract description 2
- 238000013329 compounding Methods 0.000 abstract 1
- 230000009982 effect on human Effects 0.000 abstract 1
- 210000002149 gonad Anatomy 0.000 abstract 1
- 239000007788 liquid Substances 0.000 abstract 1
- 238000005516 engineering process Methods 0.000 description 7
- 102000016911 Deoxyribonucleases Human genes 0.000 description 4
- 108010053770 Deoxyribonucleases Proteins 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 238000001311 chemical methods and process Methods 0.000 description 3
- 239000013505 freshwater Substances 0.000 description 3
- 238000013016 damping Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 230000002710 gonadal effect Effects 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 241000276401 Batrachoididae gen. sp. Species 0.000 description 1
- 241000252233 Cyprinus carpio Species 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 208000002193 Pain Diseases 0.000 description 1
- 108010059712 Pronase Proteins 0.000 description 1
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 1
- 241000269841 Thunnus albacares Species 0.000 description 1
- 210000004712 air sac Anatomy 0.000 description 1
- 230000003796 beauty Effects 0.000 description 1
- 238000002306 biochemical method Methods 0.000 description 1
- 244000309466 calf Species 0.000 description 1
- 241001233037 catfish Species 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 238000005360 mashing Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 230000036407 pain Effects 0.000 description 1
- XXPDBLUZJRXNNZ-UHFFFAOYSA-N promethazine hydrochloride Chemical compound Cl.C1=CC=C2N(CC(C)N(C)C)C3=CC=CC=C3SC2=C1 XXPDBLUZJRXNNZ-UHFFFAOYSA-N 0.000 description 1
- 239000013535 sea water Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 210000004336 spermatogonium Anatomy 0.000 description 1
- 210000001550 testis Anatomy 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000009333 weeding Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Landscapes
- Meat, Egg Or Seafood Products (AREA)
Abstract
The invention relates to a process for extracting deoxyribonucleic acid (DNA), and provides a production process for extracting DNA from gonads of fishes such as spermary of the fishes. The technological process includes compounding mixed liquid of sodium citrate and sodium chloride, adding salt and sodium bicarbonate as production reagent, successively adding fish spermary, stirring, centrifuging to leave precipitate, and treating with pancreatin and edible alcohol to obtain white or yellowish fibrous product. The invention has simple process, time and material saving, high extraction rate, no environmental pollution, no toxic or side effect on human body and wide application range.
Description
The present invention relates to the extraction process of thymus nucleic acid, further relate to the technology of extracting thymus nucleic acid (DNA) from piscinity gland such as spermary.
Thymus nucleic acid (Deoxytihonucleic acid-writes a Chinese character in simplified form DNA) is the main genetic material of human body and organism, and it plays control action kou to vital movements such as the growth of biology, growth, heredity, variation, metabolism.
The thymus nucleic acid product, spectrum of use is extremely extensively removed as biochemical reagents, outside the raw materials such as extraction Nucleotide, in medicines and health protection, aspects such as agricultural crops and food beauty treatment all produce effect quite significantly, therefore all classify domestic and international major research item always as, attract investigators' ground ploughing and weeding of sparing no pains.
Earlier 1900s, the researchist can only adopt chemical process under lab to carry out trace and extract from the spermary of toadfish and catfish, has found again after a while and can extract from the thymus gland of calf; Invented the technology of extracting DNA from the albacore spermary at present both at home and abroad in succession, these technology are finished by chemical process mostly, yet the drawback of chemical process is self-evident.
Russian patent SU652187 and Japanese Patent J55015449 have disclosed the technology of extracting DNA in the gonadal tissue body of fish in succession, the former usefulness be biochemical extraction method, what the latter used is the chemical extraction method.Yet the biochemical extraction method of the former usefulness is to adopt trypsinase or PRONASE A to handle, the cost height, the source is few, and complex procedures, can only in the laboratory, extract on a small quantity, do not form scale operation, moreover extract DNA with biochemical method from the fresh water carp testis, settled the present does not still have report both at home and abroad.
The objective of the invention is to:
1, enlarges the industrial scale of DNA, extensively draw materials, extract it from gonadal tissue such as the fish spermary of all kinds of fishes, and then keep away the drawback of chemical extraction method, raise the advantage of biochemical extraction method, reduce environmental pollution.
2, simplify technology, shorten the production time, improve extraction yield.
Technical scheme of the present invention is as follows:
Get fish spermary (fresh water sea water fish all can) and make raw material, because of the spermatogonium in the spermary contains a large amount of DNA, spermary itself contains deoxyribonuclease (DNase) again, adds Trisodium Citrate in the course of processing, the activity that suppresses DNase makes DNA in the spermary not by the hydrolysis of the DNase of itself institute.Adopt enzymatic protein, make it to break away from, about 30 ℃, add the NaHCO of PH9 with DNA
3Damping fluid is used edible ethanol deposit D NA at last, makes DNA become fibrous precipitation, and after filtration, drying obtains white or little yellow fibers shape DNA Industrial products.
Below in conjunction with technique scheme in detail the present invention is described in detail:
The present invention adopts the mixed solution (hereinafter to be referred as SSC) of Trisodium Citrate (Sodium-Citrate) and sodium-chlor (Sodium Clrororide), sodium bicarbonate (NaHCO
3) and purity be that 95% edible industrial spirit is a reaction reagent.Trisodium Citrate in the described SSC solution is a food grade, gets 150g and is dissolved in the 10000ml water and stirs, and makes it dissolving, adds 60g salt (NaCl) again and adds sodium citrate solution, stirs and makes it dissolving.Get 100g food grade sodium bicarbonate (NaHCO
3) add in the 10000ml water, make it dissolving, the NaHCO of PH9
3Damping fluid.
Technical process of the present invention is specially 7 steps.
1, the inclusion in the spermary (as fat, clot, liver, air bladder, reticular tissue etc.) is removed totally, and, after draining away the water, weighed with the clear water washing.
2, the spermary of weighing is poured into the double SSC solution of weight of tissue mashing machine, smash to pieces and make into homogenate.
3, homogenate is poured in the centrifuge tube, balance on balance was changeed in/1 minute the refrigerated centrifuge centrifugal 8-11 minute 3000 again, and the supernatant liquor that inclines stays precipitation.
4, above operation triplicate, purpose are to remove water-soluble and the salt solubility impurity, stay precipitation.
5, will precipitate taking-up, add the PH9 sodium hydrogen carbonate solution of suitable raw material weight 1/20, and add and mix pancreatin 0.08-0.1%, stir, what is incubated 5-10 minute down for 37 ℃, and summer, room temperature got final product, continue to stir, pour 95% alcohol after hydrolysis finishes into and make the DNA precipitation.
6, repeatedly with filter dry wine essence after the alcohol precipitation dehydration three times, get product.
7, put in the air dry oven about 60 ℃ white or little yellow fibers shape finished product dry.
Compared with the prior art, advantage of the present invention is embodied in the following aspects:
1, production technology is simple, only needs 2-2.5 hour from being dosed into out finished product, and feed consumption is few, and yield is big, and average recovery rate is more than 10%, and technology will be spent 2-3 days before, and recovery rate is 4-5% only.
2, no environmental pollution, the working condition gentleness, used reagent is edible level, human body is not had secondary toxic action, and the in the past deproteinized agent of usefulness is dodecyl sodium sulfate or organic solvent, expends big and the pollution environment, and the product of producing can not be used for food additives.
3, draw materials easily, wide material sources except extracting from the fish spermary, also can extract from the fresh-water fishes spermary.
Claims (4)
1, thymus nucleic acid of the present invention (DNA) extraction process is through operations such as (homogenate, precipitation, stirrings), it is characterized in that the present invention is the spermary impurity elimination with fish, clean and drain the mixed solution that the back adds Trisodium Citrate and sodium-chlor, smash to pieces, homogenate, pour centrifuge tube then into, balance on the what balance, changeed in/1 minute the refrigerated centrifuge centrifugal 8-11 minute 3000 again, supernatant liquor inclines, stay precipitation, above-mentioned operation triplicate takes out precipitation then, is incorporated as the sodium bicarbonate (NaHCO of raw material weight 1/20
3) the pancreas mixed enzyme of solution and 0.08-0.1%, stirring, 35 ℃-37.5 ℃ insulations of what 5-10 minute stir, and hydrolysis finishes, and pours 95% alcohol precipitation into, and the smart drying of filter dry wine gets product.
2, according to the described thymus nucleic acid of claim 1 (DNA) extraction process, it is characterized in that sodium bicarbonate (NaHCO
3) solution acid alkalinity is PH9.
3,, it is characterized in that with 95% alcohol repeated precipitation white fiber shape finished product three times according to the described thymus nucleic acid of claim 1 (DNA) extraction process.
4,, it is characterized in that described Trisodium Citrate and sodium bicarbonate, alcohol are food grade according to the described thymus nucleic acid of claim 1 (DNA) extraction process.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 94110993 CN1112124A (en) | 1994-05-16 | 1994-05-16 | Process for extracting deoxyribonucleic acid (DNA) |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 94110993 CN1112124A (en) | 1994-05-16 | 1994-05-16 | Process for extracting deoxyribonucleic acid (DNA) |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1112124A true CN1112124A (en) | 1995-11-22 |
Family
ID=5034905
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 94110993 Pending CN1112124A (en) | 1994-05-16 | 1994-05-16 | Process for extracting deoxyribonucleic acid (DNA) |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1112124A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1052728C (en) * | 1997-05-05 | 2000-05-24 | 四川师范大学 | Method for extracting hologenome of giant panda |
| CN110760509A (en) * | 2019-07-03 | 2020-02-07 | 深圳瑞达生物股份有限公司 | Method for extracting milt component of globefish testis |
| CN112341503A (en) * | 2020-11-07 | 2021-02-09 | 潍坊华诺医药科技有限公司 | Separation and purification method of deoxyribonucleoside triphosphate |
| CN113416728A (en) * | 2021-07-15 | 2021-09-21 | 中国海洋大学 | Method for extracting edible milt DNA |
-
1994
- 1994-05-16 CN CN 94110993 patent/CN1112124A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1052728C (en) * | 1997-05-05 | 2000-05-24 | 四川师范大学 | Method for extracting hologenome of giant panda |
| CN110760509A (en) * | 2019-07-03 | 2020-02-07 | 深圳瑞达生物股份有限公司 | Method for extracting milt component of globefish testis |
| CN112341503A (en) * | 2020-11-07 | 2021-02-09 | 潍坊华诺医药科技有限公司 | Separation and purification method of deoxyribonucleoside triphosphate |
| CN112341503B (en) * | 2020-11-07 | 2022-05-20 | 潍坊华诺医药科技有限公司 | Method for separating and purifying deoxyribonucleoside triphosphate |
| CN113416728A (en) * | 2021-07-15 | 2021-09-21 | 中国海洋大学 | Method for extracting edible milt DNA |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C01 | Deemed withdrawal of patent application (patent law 1993) | ||
| WD01 | Invention patent application deemed withdrawn after publication |