CN104585046A - Large-scale preparation method of MT (carbon source-contained) solid culture medium - Google Patents
Large-scale preparation method of MT (carbon source-contained) solid culture medium Download PDFInfo
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- CN104585046A CN104585046A CN201510082411.4A CN201510082411A CN104585046A CN 104585046 A CN104585046 A CN 104585046A CN 201510082411 A CN201510082411 A CN 201510082411A CN 104585046 A CN104585046 A CN 104585046A
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Abstract
The invention discloses a large-scale preparation method of an MT (carbon source-contained) solid culture medium, and belongs to the technical field of plant biologics. The large-scale preparation method comprises the following steps: firstly, calculating the demand amounts of 17 elements of the MT (carbon source-contained) solid culture medium after being amplified for 1000 times; accurately weighing the amplified amounts of medicines; sequentially uniformly mixing the 17 elements according to the demand amounts, and sealing the MT (carbon source-contained) solid culture medium; unsealing to use the solid culture medium according to demands. The method can prepare the MT (carbon source-contained) solid culture medium, has the characteristics of being convenient and rapid to operate, simple to use and small in errors of different batches and same batches, prevents the phenomena of large errors, deposition, mildew and the like caused by artificial operation and external factors, and can be applied to biological technical purposes such as plant tissue culture.
Description
Technical field
The invention discloses the scale preparation method of a kind of MT (carbonaceous sources) solid culture matrix, belong to plant biotechnology field.Be more particularly one include accurate calculating, accurate weighing, first few after how mix, seal, the five steps such as use of breaking seal, to obtain different batches and batch between the minimum culture matrix of error, minimizing experimental error method.
Background technology
Plant tissue culture technique or cultivation technique without soil need preparation medium mother liquor usually, and namely preparation expands certain multiple, liquid containing all kinds reagent.But, because mother liquor contains dissimilar reagent, be usually divided into the large class of macroelement, trace element, molysite and organic element four, and the multiple that four large classes expand is different, causes medicine service property (quality) inconsistent, easily occurs the error of calculation first; Second the producer of medicine, quality are inconsistent, even if to calculate, weigh consistent, also can affect result of the test; Three due to mother liquor be liquid condition, at the end of spring and the beginning of summer season, because the temperature rises and humidity increases, extremely easily cause mother liquor bacterial infection, mould, cause the phenomenon presenting different colours in mother liquor or occur precipitation, waste raw material; Comparatively big error can be there is to the difference of the cognition of container scale (especially trace element), instrument precision in four fundamental rules because of weighing person.
In view of the above-mentioned shortcoming of mother liquor; with the mother liquor medium of routine; usually there will be the failed phenomenon that maybe cannot repeat of the result of the test caused because of factors such as weighing person, medicine, seasons; have impact on the process using Plant Tissue Breeding or cultivation technique without soil to carry out test greatly, hit the confidence of experimenter.
Summary of the invention
A kind of scale preparation method of MT (carbonaceous sources) solid culture matrix, is characterized in that, include in described method accurate calculating, accurate weighing, first few after how mix, seal, the five steps such as use of breaking seal.
Method according to claim 1, is characterized in that, described five steps concrete operations are as follows:
1. accurately calculate: 17 kinds of elements contained by MT (carbonaceous sources) solid-based are expanded 1000 times, i.e. CoCL2 6H2O and each 0.025g of cupric sulfate pentahydrate, nicotinic acid 0.5g, puridoxine hydrochloride 0.5g, potassium iodide 0.83g, glycine 2g, boric acid 6.2g, white vitriol 8.6g, four water manganese sulphate 22.3g, iron edetate 36.5g, inositol 100g, potassium dihydrogen phosphate 170g, epsom salt 370g, calcium chloride dihydrate 440g, ammonium nitrate 1650g, potassium nitrate 1900g, soft white sugar 30000g;
2. accurate weighing: according to the result precise of 1. middle calculating, especially with CoCL2 6H2O and the cupric sulfate pentahydrate of each 0.025g of ten thousand/balance precise;
3. how to mix after first few: mix according to few priority of carrying out of amount more, namely according to CoCL2 6H2O and cupric sulfate pentahydrate, nicotinic acid, puridoxine hydrochloride, potassium iodide, glycine, boric acid, white vitriol, four water manganese sulphates, iron edetate, inositol, potassium dihydrogen phosphate, epsom salt, calcium chloride dihydrate, ammonium nitrate, potassium nitrate, the order of soft white sugar mixes, and amounts to 34707.48g;
4. seal: the requirement of mixed solid matrix according to 250g every bottle is sub-packed in plastic containers, and seals;
5. to break seal use: as required, add the above-mentioned matrix 34.7g of precise in every 1L distilled water, heating, after it dissolves completely, adds reagent needed for other.
Beneficial effect of the present invention: without the error of calculation between the matrix of different batches; Because all reagent all expands 1000 times, the weighing error between the matrix of different batches is minimum; In theory, owing to being the usage amount (calculating with each 1L) of 1000 times at every turn, namely can prepare the above-mentioned formula of 1000L, batch inside there will not be the human error caused because using the reagent of different manufacturers, different times; Matrix after expansion is that every 250g is sealed in plastic containers, can avoid because the impact of the extraneous factor such as weather, temperature, moisture causes occurring the common precipitation of liquid mother liquor, the phenomenon such as mouldy; Above-mentioned plurality of advantages is all the follow-up test carried out for matrix with this medium, obtains result of the test accurately and lays the foundation.
Embodiment:
Below in conjunction with specific embodiment, the present invention is conducted further description.
The scale preparation method of a kind of MT (carbonaceous sources) solid culture matrix in the present embodiment, include accurate calculating, accurate weighing, first few after how mix, seal, the five steps such as use of breaking seal.
Above-mentioned five steps concrete operations are as follows:
1. accurately calculate: 17 kinds of elements contained by MT (carbonaceous sources) solid-based are expanded 1000 times, i.e. CoCL2 6H2O and each 0.025g of cupric sulfate pentahydrate, nicotinic acid 0.5g, puridoxine hydrochloride 0.5g, potassium iodide 0.83g, glycine 2g, boric acid 6.2g, white vitriol 8.6g, four water manganese sulphate 22.3g, iron edetate 36.5g, inositol 100g, potassium dihydrogen phosphate 170g, epsom salt 370g, calcium chloride dihydrate 440g, ammonium nitrate 1650g, potassium nitrate 1900g, soft white sugar 30000g;
2. accurate weighing: according to the result precise of 1. middle calculating, especially with CoCL2 6H2O and the cupric sulfate pentahydrate of each 0.025g of ten thousand/balance precise;
3. how to mix after first few: mix according to few priority of carrying out of amount more, namely according to CoCL2 6H2O and cupric sulfate pentahydrate, nicotinic acid, puridoxine hydrochloride, potassium iodide, glycine, boric acid, white vitriol, four water manganese sulphates, iron edetate, inositol, potassium dihydrogen phosphate, epsom salt, calcium chloride dihydrate, ammonium nitrate, potassium nitrate, the order of soft white sugar mixes, and amounts to 34707.48g;
4. seal: the requirement of mixed solid matrix according to 250g every bottle is sub-packed in plastic containers, and seals;
5. to break seal use: as required, add the above-mentioned matrix 34.7g of precise in every 1L distilled water, heating, after it dissolves completely, adds reagent needed for other.
1L solution after above-mentioned substrate preparation is added the agar powder of 5g, and then add the KOH of 1N of 1ml, dissolve completely and be sub-packed in the vial of 30 200ml, after 121 DEG C of sterilizing 20min.On superclean bench, inoculate stevia stem section, after cultivating 30d under 25 DEG C of illumination 12h conditions, occur axillalry bud and take root good.
Claims (3)
1. a scale preparation method for MT (carbonaceous sources) solid culture matrix, is characterized in that, include in described method accurate calculating, accurate weighing, first few after how mix, seal, the five steps such as use of breaking seal.
2. method according to claim 1, is characterized in that, described five steps concrete operations are as follows:
1. accurately calculate: 17 kinds of elements contained by MT (carbonaceous sources) solid-based are expanded 1000 times, i.e. CoCL2 6H2O and each 0.025g of cupric sulfate pentahydrate, nicotinic acid 0.5g, puridoxine hydrochloride 0.5g, potassium iodide 0.83g, glycine 2g, boric acid 6.2g, white vitriol 8.6g, four water manganese sulphate 22.3g, iron edetate 36.5g, inositol 100g, potassium dihydrogen phosphate 170g, epsom salt 370g, calcium chloride dihydrate 440g, ammonium nitrate 1650g, potassium nitrate 1900g, soft white sugar 30000g;
2. accurate weighing: according to the result precise of 1. middle calculating, especially with CoCL2 6H2O and the cupric sulfate pentahydrate of each 0.025g of ten thousand/balance precise;
3. how to mix after first few: mix according to few priority of carrying out of amount more, namely according to CoCL2 6H2O and cupric sulfate pentahydrate, nicotinic acid, puridoxine hydrochloride, potassium iodide, glycine, boric acid, white vitriol, four water manganese sulphates, iron edetate, inositol, potassium dihydrogen phosphate, epsom salt, calcium chloride dihydrate, ammonium nitrate, potassium nitrate, the order of soft white sugar mixes, and amounts to 34707.48g;
4. seal: the requirement of mixed solid matrix according to 250g every bottle is sub-packed in plastic containers, and seals;
5. to break seal use: as required, add the above-mentioned matrix 34.7g of precise in every 1L distilled water, heating, after it dissolves completely, adds reagent needed for other.
3. method according to claim 1, is characterized in that, the scale preparation method of this culture matrix has following characteristics, i.e. without the error of calculation between the matrix of different batches; Because all reagent all expands 1000 times, the weighing error between the matrix of different batches is minimum; In theory, owing to being the usage amount (calculating with each 1L) of 1000 times at every turn, namely can prepare the above-mentioned formula of 1000L, batch inside there will not be the human error caused because using the reagent of different manufacturers, different times; Matrix after expansion is that every 250g is sealed in plastic containers, can avoid because the impact of the extraneous factor such as weather, temperature, moisture causes occurring the common precipitation of liquid mother liquor, the phenomenon such as mouldy; Above-mentioned plurality of advantages is all the follow-up test carried out for matrix with this medium, obtains result of the test accurately and lays the foundation.
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| CN201510082411.4A CN104585046A (en) | 2015-02-16 | 2015-02-16 | Large-scale preparation method of MT (carbon source-contained) solid culture medium |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101070531A (en) * | 2006-05-09 | 2007-11-14 | 陈曦 | Dry-powder-type culturing substrate and preparing method |
| CN101386836A (en) * | 2007-09-12 | 2009-03-18 | 北京清大天一生物技术有限公司 | Zooblast culture medium dry powder composition, culture medium composition and preparation method thereof |
| WO2014147869A1 (en) * | 2013-03-22 | 2014-09-25 | Mizuta Youichi | Method for preparing culture medium for culturing plant tissue, method for culturing plant tissue, sterilizing agent, germicidal agent, and culture medium composition for culturing plant tissue |
| CN104151001A (en) * | 2014-08-18 | 2014-11-19 | 江西省江天农业科技有限公司 | Preparation method of MS medium |
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2015
- 2015-02-16 CN CN201510082411.4A patent/CN104585046A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101070531A (en) * | 2006-05-09 | 2007-11-14 | 陈曦 | Dry-powder-type culturing substrate and preparing method |
| CN101386836A (en) * | 2007-09-12 | 2009-03-18 | 北京清大天一生物技术有限公司 | Zooblast culture medium dry powder composition, culture medium composition and preparation method thereof |
| WO2014147869A1 (en) * | 2013-03-22 | 2014-09-25 | Mizuta Youichi | Method for preparing culture medium for culturing plant tissue, method for culturing plant tissue, sterilizing agent, germicidal agent, and culture medium composition for culturing plant tissue |
| CN104151001A (en) * | 2014-08-18 | 2014-11-19 | 江西省江天农业科技有限公司 | Preparation method of MS medium |
Non-Patent Citations (1)
| Title |
|---|
| 彭正松等: "《遗传学实验教程》", 30 September 2012 * |
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Application publication date: 20150506 |