CN109380113A - A kind of proliferated culture medium manufacture craft - Google Patents
A kind of proliferated culture medium manufacture craft Download PDFInfo
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- CN109380113A CN109380113A CN201710655250.2A CN201710655250A CN109380113A CN 109380113 A CN109380113 A CN 109380113A CN 201710655250 A CN201710655250 A CN 201710655250A CN 109380113 A CN109380113 A CN 109380113A
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- Prior art keywords
- mother liquor
- preparation
- plant growth
- growth regulating
- substance
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- 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.)
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
Abstract
The present invention relates to a kind of proliferated culture medium manufacture crafts, it uses following method and step: step 1: a great number of elements mother liquor preparation；Step 2: microelement mother liquor preparation；Step 3: mother liquid of iron salt preparation；Step 4: organic principle mother liquor preparation；Step 5: the preparation of indolebutyric acid plant growth regulating substance mother liquor；Or the preparation of α-naphthylacetic acid plant growth regulating substance mother liquor；Step 6: 6- benzyl purine plant growth regulating substance mother liquor preparation；Step 7: being put into some distilled water in beaker, weighs 7g or so agar powder, is placed on and is heated to boiling on electric furnace, until agar powder dissolves；Step 8: weighing 30g or so sucrose, and stirring and dissolving in agar solution is added, and forms mixed liquor；The advantages that it uses a variety of mother liquors, has preparation process simple, at low cost, can improve the growth coefficient, growth quality and culture efficiency of plant tissue.
Description
[technical field]
The present invention relates to a kind of proliferated culture medium manufacture crafts for the degeneration-resistant excellent dwarfing rootstock KM23 of apple.
[background technique]
China is a large agricultural country, and as people's lives level improves, demand of the people for all kinds of fruit
It is higher and higher, therefore fruit growing gradually grows up.Stock is the basis of fruit tree growth, for controlling tree body growing way, increases fruit
Drought resisting, cold-resistant, salt resistance alkali, resistant to diseases and insects are set, expands fruit variety adaptation range and is of great significance.And the growth of fruit tree
Cheng Zhong, serving for proliferated culture medium are larger.Proliferated culture medium is in fermentation process or animal and plant cells mass propgation for micro- life
Growth, breeding or the accumulation metabolite of object or animal and plant cell, with nutrient matrix necessary to synthesising biological chemical products.
The complex manufacturing technology of traditional proliferated culture medium, higher cost, and can not accomplish accurately to control, eventually lead to quality decline.
[summary of the invention]
In view of the defects and deficiencies of the prior art, the present invention intends to provide a kind of proliferated culture medium manufacture craft,
It is customized using a variety of mother liquors, is had preparation process simple, at low cost, can be improved growth coefficient, the growth quality of plant tissue,
And culture efficiency, it has high economic benefit.
A kind of proliferated culture medium manufacture craft of the present invention, it is adopted the following technical scheme that:
Step 1: the preparation of a great number of elements mother liquor: it uses following method and step:
1) potassium nitrate KNO is taken3190g/L, ammonium nitrate NH4NO3165g/L, potassium dihydrogen phosphate KH2PO417g/L, magnesium sulfate
MgsO4.7H2O 37g/L, calcium chloride CaCl2.2H2O 44g/L；
2) by above-mentioned steps 1) each substance be respectively placed in stirring cup, distilled water is added and thoroughly dissolves, then mixing is fixed
Hold 1L, forms a great number of elements mother liquor stock solution；
Step 2: the preparation of microelement mother liquor: it uses following method and step:
1) potassium iodide KI 0.083g/L, boric acid H are taken3BO30.62g/L, manganese sulfate MnSO4.4H2O 2.23g/L, sulfuric acid
Zinc ZnSO4.7 H2O 0.86g/L, sodium molybdate Na2MoO4.2 H2O 0.025g/L, copper sulphate CuSO4.5 H2O 0.0025g/L,
Cobalt chloride CoCl20.0025g/L:
2) each substance in step 1) is respectively placed in stirring cup, distilled water is added and thoroughly dissolves, then mixes constant volume
To 1L, microelement mother liquor stock solution is formed；
Step 3: mother liquid of iron salt preparation: it uses following method and step:
1) disodium ethylene diamine tetraacetate Na is taken2.EDTA 3.73g/L, ferrous sulfate FeSO4.7H2O 2.78g/L:
2) each substance in step 1) is respectively placed in stirring cup, distilled water is added and thoroughly dissolves, then mixes constant volume
To 1L, mother liquid of iron salt stock solution is formed；
Step 4: the preparation of organic principle mother liquor: it uses following method and step:
1) inositol 10g/L, glycine 0.2g/L, thiamine hydrochloride VB1 0.01g/L, puridoxine hydrochloride VB6 0.05g/ are taken
L, niacin VB5 0.05g/L；
2) each substance in step 1) is respectively placed in stirring cup, distilled water is added and thoroughly dissolves, then mixes constant volume
To 1L, organic principle mother liquor stock solution is formed；
Step 5: the preparation of indolebutyric acid plant growth regulating substance mother liquor: its step with the following method:
1) indolebutyric acid 10mg is weighed；
2) indolebutyric acid is dissolved with the HCL solution that the substance withdrawl syndrome of 1mL is 0.1mol/L, course of dissolution needs water
Bath heating, is finally settled to 100mL, obtains the indolebutyric acid plant growth regulating substance mother liquor that mass concentration is 0.1mg/mL；
Or the preparation of α-naphthylacetic acid plant growth regulating substance mother liquor: its step with the following method:
1) α-naphthylacetic acid 10mg is weighed；
2) a small amount of 75% alcoholic solution of α-naphthylacetic acid is dissolved, is finally settled to 100mL, obtaining mass concentration is 0.1mg/
The α-naphthylacetic acid plant growth regulating substance mother liquor of mL；
Step 6: the preparation of 6- benzyl purine plant growth regulating substance mother liquor: it uses following method and step:
1) 6- benzyl purine 10mg is weighed；
2) 6- benzyl purine is dissolved with the NaOH solution that the substance withdrawl syndrome of 1mL is 0.1mol/L, course of dissolution needs
Heating water bath is wanted, 100mL is finally settled to, it is female to obtain the 6- benzyl purine plant growth regulating substance that mass concentration is 0.1mg/mL
Liquid；
Step 7: being put into some distilled water in beaker, weighs 7g or so agar powder, is placed on and is heated to boiling on electric furnace,
Until agar powder dissolves；
Step 8: using graduated cylinder or pipette, takes a great number of elements mother liquor stock solution 1-15ml in step 1, in step 2
Organic principle in mother liquid of iron salt stock solution 1-15ml, step 4 in microelement mother liquor stock solution 1-15ml, step 3 is female
Indolebutyric acid plant growth regulating substance mother liquor 0.01-5ml in liquid stock solution 1-15ml, step 5 or α-naphthalene in step 5
6- benzyl purine plant growth regulating substance mother liquor 1-8ml in acetic acid plant growth regulating substance mother liquor 0.1-5ml, step 6,
It pours into beaker；
Step 9: weighing 30g sucrose and pour into beaker, stirring and dissolving, forms mixed liquor；
Step 10: distilled water will be added in beaker, mixed liquor is then settled to 1L；
Step 11: being measured with precision test paper or acidometer, adjusts PH to 5.7- with the NaOH or 1mol/LHCL of 1mol/L
5.8；
Step 12: it after slightly cooling down, is distributed into culture vessel, the culture vessel sealed membrane or brown paper of uncovered
Sealing, is tightened with rubber band or rope；
Step 13: the culture vessel in upper step 12 is put into the sterilizing of sterilization pot, 121 DEG C of sterilizings, 20 minutes left sides
It is right；
Step 14: taking out culture medium from autoclave after sterilizing, at room temperature after natural cooling, forms finished product culture
Base.
After adopting the above structure, the invention has the following beneficial effects: a kind of proliferated culture medium manufacture craft of the present invention, it
It is customized using a variety of mother liquors, has preparation process simple, at low cost, growth coefficient, the growth quality of plant tissue can be improved, with
And culture efficiency, it has high economic benefit.
[specific embodiment]
With specific embodiment, the present invention will be described in detail below, and illustrative examples therein and explanation are only used to explain
The present invention, but it is not as a limitation of the invention.
A kind of proliferated culture medium manufacture craft described in present embodiment, it is adopted the following technical scheme that:
Step 1: the preparation of a great number of elements mother liquor: it uses following method and step:
1) potassium nitrate KNO is taken3190g/L, ammonium nitrate NH4NO3165g/L, potassium dihydrogen phosphate KH2PO417g/L, magnesium sulfate
MgSO4.7H2O 37g/L, calcium chloride CaCl2.2H2O 44g/L；
2) by above-mentioned steps 1) each substance be respectively placed in stirring cup, distilled water is added and thoroughly dissolves, then mixing is fixed
Hold 1L, forms a great number of elements mother liquor stock solution；
Step 2: the preparation of microelement mother liquor: it uses following method and step:
1) potassium iodide KI 0.083g/L, boric acid H are taken3BO30.62g/L, manganese sulfate MnSO4.4H2O 2.23g/L, sulfuric acid
Zinc ZnSO4.7 H2O 0.86g/L, sodium molybdate Na2MoO4.2 H2O 0.025g/L, copper sulphate CuSO4.5 H2O 0.0025g/L,
Cobalt chloride CoCl20.0025g/L；
2) each substance in step 1) is respectively placed in stirring cup, distilled water is added and thoroughly dissolves, then mixes constant volume
To 1L, microelement mother liquor stock solution is formed；
Step 3: mother liquid of iron salt preparation: it uses following method and step:
1) disodium ethylene diamine tetraacetate Na is taken2.EDTA 3.73g/L, ferrous sulfate FeSO4.7H2O 2.78g/L；
2) each substance in step 1) is respectively placed in stirring cup, distilled water is added and thoroughly dissolves, then mixes constant volume
To 1L, mother liquid of iron salt stock solution is formed；
Step 4: the preparation of organic principle mother liquor: it uses following method and step:
1) inositol 10g/L, glycine 0.2g/L, thiamine hydrochloride VB1 0.01g/L, puridoxine hydrochloride VB6 0.05g/ are taken
L, niacin VB5 0.05g/L；
2) each substance in step 1) is respectively placed in stirring cup, distilled water is added and thoroughly dissolves, then mixes constant volume
To 1L, organic principle mother liquor stock solution is formed；
Step 5: the preparation of indolebutyric acid plant growth regulating substance mother liquor: its step with the following method:
1) indolebutyric acid 10mg is weighed；
2) indolebutyric acid is dissolved with the HCL solution that the substance withdrawl syndrome of 1mL is 0.1mol/L, course of dissolution needs water
Bath heating, is finally settled to 100mL, obtains the indolebutyric acid plant growth regulating substance mother liquor that mass concentration is 0.1mg/mL；
Or the preparation of α-naphthylacetic acid plant growth regulating substance mother liquor: its step with the following method:
1) α-naphthylacetic acid 10mg is weighed；
2) a small amount of 75% alcoholic solution of α-naphthylacetic acid is dissolved, is finally settled to 100mL, obtaining mass concentration is 0.1mg/
The α-naphthylacetic acid plant growth regulating substance mother liquor of mL；
Step 6: the preparation of 6- benzyl purine plant growth regulating substance mother liquor: it uses following method and step:
1) 6- benzyl purine 10mg is weighed；
2) 6- benzyl purine is dissolved with the NaOH solution that the substance withdrawl syndrome of 1mL is 0.1mol/L, course of dissolution needs
Heating water bath is wanted, 100mL is finally settled to, it is female to obtain the 6- benzyl purine plant growth regulating substance that mass concentration is 0.1mg/mL
Liquid；
Step 7: being put into some distilled water in beaker, weighs 7g or so agar powder, is placed on and is heated to boiling on electric furnace,
Until agar powder dissolves；
Step 8: using graduated cylinder or pipette, takes a great number of elements mother liquor stock solution 1-15ml in step 1, in step 2
Organic principle in mother liquid of iron salt stock solution 1-15ml, step 4 in microelement mother liquor stock solution 1-15ml, step 3 is female
Indolebutyric acid plant growth regulating substance mother liquor 0.01-5ml in liquid stock solution 1-15ml, step 5 or α-naphthalene in step 5
6- benzyl purine plant growth regulating substance mother liquor 1-8ml in acetic acid plant growth regulating substance mother liquor 0.1-5ml, step 6,
It pours into beaker；
Step 9: weighing 30g sucrose and pour into beaker, stirring and dissolving, forms mixed liquor；
Step 10: distilled water will be added in beaker, mixed liquor is then settled to 1L；
Step 11: being measured with precision test paper or acidometer, adjusts PH to 5.7- with the NaOH or 1mol/LHCL of 1mol/L
5.8；
Step 12: it after slightly cooling down, is distributed into culture vessel, the culture vessel sealed membrane or brown paper of uncovered
Sealing, is tightened with rubber band or rope；
Step 13: the culture vessel in upper step 12 is put into the sterilizing of sterilization pot, 121 DEG C of sterilizings, 20 minutes left sides
It is right；
Step 14: taking out culture medium from autoclave after sterilizing, at room temperature after natural cooling, forms finished product culture
Base.
In the present invention, Ms preparation of culture medium:
1) a great number of elements: potassium nitrate KNO3190g/L, ammonium nitrate NH4NO3165g/L, potassium dihydrogen phosphate KH2PO4 17g/
L, magnesium sulfate MgSO4.7H2O 37g/L, calcium chloride CaCl2.2H2O 44g/L；
2) microelement: potassium iodide KI 0.083g/L, boric acid H3BO30.62g/L, manganese sulfate MnSO4.4H2O 2.23g/
L, zinc sulfate ZnSO4.7 H2O 0.86g/L, sodium molybdate Na2MoO4.2 H2O 0.025g/L, copper sulphate CuSO4.5 H2O
0.0025g/L, cobalt chloride CoCl20.0025g/L；
3) molysite: disodium ethylene diamine tetraacetate Na2.EDTA 3.73g/L, ferrous sulfate eSO4.7H2O2.78g/L；
4) organic principle: inositol 10g/L, glycine 0.2g/L, thiamine hydrochloride VB1 0.01g/L, puridoxine hydrochloride VB6
0.05g/L, niacin VB5 0.05g/L；
5) sucrose sucrose 30g/L, agar agar 7g/L.
6) above-mentioned several mother liquors will individually be made into the stock solution of 1L.Its preparation method is: in every kind of mother liquor it is several at
Divide after weighing, is thoroughly dissolved with a small amount of distilled water respectively, then again that they are miscible, last constant volume to 1L.
It also needs to be added in Ms culture medium: indolebutyric acid (abbreviation IBA) or α-naphthylacetic acid (abbreviation NAA), 6- benzyl purine
Plant growth regulating substances such as (abbreviation 6-BA), and it is made into mother liquor 0.1mg/mL respectively.Its preparation method is: weighing this respectively
3 kinds of each 10mg of substance dissolve IBA with the HCL solution that the substance withdrawl syndrome of a small amount of 1mL is 0.1mol/L, by 6-BA with less
The NaOH solution that the substance withdrawl syndrome for measuring 1mL is 0.1mol/L dissolves, and a small amount of 75% essence of NAA is dissolved, finally fixed respectively
Hold the mother liquor for being 0.1mg/mL to get mass concentration to 100mL.
Prepare culture solution: take out required dosage respectively from various mother liquors with graduated cylinder or pipette: Ms mother liquor is 1-
15mL, then 6-BA1-8mL, IBA 0.01-5ml or NAA0.1-5ml are taken, it is put into togerther in beaker with various mother liquors.
In the present invention, for configuring 1L Ms culture medium, proceed as follows in order, operating process is as follows:
1) it is first put into some distilled water in beaker, weighs 7g or so agar powder, is placed on and is heated to boiling on electric furnace, until
Agar powder dissolves.
2) various mother liquors are taken out by formula to pour into.
3) it generally weighs 30g sucrose to pour into, stirring and dissolving.
4) plus distilled water is settled to 1L.
5) it is measured with precision test paper or acidometer, adjusts PH to 5.7-5.8 with the NaOH or 1mol/LHCL of 1mol/L.
6) it after slightly cooling down, is distributed into culture vessel, culture vessel sealed membrane or the brown paper sealing of uncovered are used
Rubber band or rope tighten.
7) it is put into the sterilizing of sterilization pot, 121 DEG C sterilize 20 minutes or so.
8) culture medium is taken out from autoclave after sterilizing, and at room temperature after natural cooling, is deposited at shady and cool dried and clean
For use.
A kind of proliferated culture medium manufacture craft of the present invention, it is customized using a variety of mother liquors, has preparation process letter
It is single, at low cost, the growth coefficient, growth quality and culture efficiency of plant tissue can be improved, is had high economic benefit.
The above description is only a preferred embodiment of the present invention, thus it is all according to feature described in present patent application range and
The equivalent change or modification that principle is done, is included in the scope of the patent application of the present invention.
Claims (1)
1. a kind of proliferated culture medium manufacture craft, it is characterised in that: it is adopted the following technical scheme that:
Step 1: the preparation of a great number of elements mother liquor: it uses following method and step:
1) potassium nitrate KNO is taken3190g/L, ammonium nitrate NH4NO3165g/L, potassium dihydrogen phosphate KH2PO417g/L, magnesium sulfate
MgSO4.7H2O 37g/L, calcium chloride CaCl2.2H2O 44g/L；
2) by above-mentioned steps 1) each substance be respectively placed in stirring cup, be added distilled water thoroughly dissolve, then mix constant volume arrive
1L forms a great number of elements mother liquor stock solution；
Step 2: the preparation of microelement mother liquor: it uses following method and step:
1) potassium iodide KI 0.083g/L, boric acid H are taken3BO30.62g/L, manganese sulfate MnSO4.4H2O 2.23g/L, zinc sulfate
ZnSO4.7H2O 0.86g/L, sodium molybdate Na2MoO4.2H2O 0.025g/L, copper sulphate CuSO4.5H2O 0.0025g/L, chlorination
Cobalt CoCl20.0025g/L；
2) each substance in step 1) is respectively placed in stirring cup, distilled water is added and thoroughly dissolves, then mixing constant volume to 1L,
Form microelement mother liquor stock solution；
Step 3: mother liquid of iron salt preparation: it uses following method and step:
1) disodium ethylene diamine tetraacetate Na is taken2.EDTA 3.73g/L, ferrous sulfate FeSO4.7H2O 2.78g/L；
2) each substance in step 1) is respectively placed in stirring cup, distilled water is added and thoroughly dissolves, then mixing constant volume to 1L,
Form mother liquid of iron salt stock solution；
Step 4: the preparation of organic principle mother liquor: it uses following method and step:
1) inositol 10g/L, glycine 0.2g/L, thiamine hydrochloride VB1 0.01g/L, puridoxine hydrochloride VB6 0.05g/L, cigarette are taken
Sour VB5 0.05g/L；
2) each substance in step 1) is respectively placed in stirring cup, distilled water is added and thoroughly dissolves, then mixing constant volume to 1L,
Form organic principle mother liquor stock solution；
Step 5: the preparation of indolebutyric acid plant growth regulating substance mother liquor: its step with the following method:
1) indolebutyric acid 10mg is weighed；
2) indolebutyric acid is dissolved with the HCL solution that the substance withdrawl syndrome of 1mL is 0.1mol/L, course of dissolution needs water-bath to add
Heat is finally settled to 100mL, obtains the indolebutyric acid plant growth regulating substance mother liquor that mass concentration is 0.1mg/mL；
Or the preparation of α-naphthylacetic acid plant growth regulating substance mother liquor: its step with the following method:
1) α-naphthylacetic acid 10mg is weighed；
2) a small amount of 75% alcoholic solution of α-naphthylacetic acid is dissolved, is finally settled to 100mL, obtaining mass concentration is 0.1mg/mL's
α-naphthylacetic acid plant growth regulating substance mother liquor；
Step 6: the preparation of 6- benzyl purine plant growth regulating substance mother liquor: it uses following method and step:
1) 6- benzyl purine 10mg is weighed；
2) 6- benzyl purine is dissolved with the NaOH solution that the substance withdrawl syndrome of 1mL is 0.1mol/L, course of dissolution needs water
Bath heating, is finally settled to 100mL, obtains the 6- benzyl purine plant growth regulating substance mother liquor that mass concentration is 0.1mg/mL；
Step 7: being put into some distilled water in beaker, weighs 7g or so agar powder, is placed on and is heated to boiling on electric furnace, until
Agar powder dissolves；
Step 8: using graduated cylinder or pipette, takes a great number of elements mother liquor stock solution 1-15ml in step 1, micro in step 2
Organic principle mother liquor storage in mother liquid of iron salt stock solution 1-15ml, step 4 in element mother liquor stock solution 1-15ml, step 3
Standby liquid 1-15ml, the indolebutyric acid plant growth regulating substance mother liquor 0.01-5ml in step 5 or the α-naphthylacetic acid in step 5
6- benzyl purine plant growth regulating substance mother liquor 1-8ml, is poured into plant growth regulating substance mother liquor 0.1-5ml, step 6
In beaker；
Step 9: weighing 30g sucrose and pour into beaker, stirring and dissolving, forms mixed liquor；
Step 10: distilled water will be added in beaker, mixed liquor is then settled to 1L；
Step 11: being measured with precision test paper or acidometer, adjusts PH to 5.7-5.8 with the NaOH or 1mol/LHCL of 1mol/L；
Step 12: after slightly cooling down, being distributed into culture vessel, culture vessel sealed membrane or the brown paper envelope of uncovered
Mouthful, it is tightened with rubber band or rope；
Step 13: the culture vessel in upper step 12 is put into the sterilizing of sterilization pot, and 121 DEG C sterilize 20 minutes or so；
Step 14: taking out culture medium from autoclave after sterilizing, at room temperature after natural cooling, forms finished product culture medium.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710655250.2A CN109380113A (en) | 2017-08-03 | 2017-08-03 | A kind of proliferated culture medium manufacture craft |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710655250.2A CN109380113A (en) | 2017-08-03 | 2017-08-03 | A kind of proliferated culture medium manufacture craft |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109380113A true CN109380113A (en) | 2019-02-26 |
Family
ID=65412370
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710655250.2A Withdrawn CN109380113A (en) | 2017-08-03 | 2017-08-03 | A kind of proliferated culture medium manufacture craft |
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| Country | Link |
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| CN (1) | CN109380113A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113998653A (en) * | 2021-09-07 | 2022-02-01 | 广西壮族自治区农业科学院 | Culture medium subpackaging device and preparation method thereof |
-
2017
- 2017-08-03 CN CN201710655250.2A patent/CN109380113A/en not_active Withdrawn
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113998653A (en) * | 2021-09-07 | 2022-02-01 | 广西壮族自治区农业科学院 | Culture medium subpackaging device and preparation method thereof |
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Application publication date: 20190226 |
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