CN104604713A - Large-scale preparation method of improved White (containing fixed media) solid culture medium - Google Patents
Large-scale preparation method of improved White (containing fixed media) solid culture medium Download PDFInfo
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- CN104604713A CN104604713A CN201510082346.5A CN201510082346A CN104604713A CN 104604713 A CN104604713 A CN 104604713A CN 201510082346 A CN201510082346 A CN 201510082346A CN 104604713 A CN104604713 A CN 104604713A
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Abstract
The invention discloses a large-scale preparation method of an improved White (containing fixed media) solid culture medium, belonging to the plant biotechnology field. The large-scale preparation method comprises the following steps: firstly computing the quantity demands after 19 elements contained in the improved White (containing fixed media) culture medium are increased by 1000 times; accurately weighing chemicals according to the chemical quantities after increase; successively mixing the 19 elements uniformly according to the quantity demands and sealing a solid medium; and unsealing the solid medium as needed to be used. The large-scale preparation method has the beneficial effects that the medium prepared by the method has the characteristics of convenience and quickness in operation, simplicity in application and small errors between different batches and in the batches; the phenomena such as large errors, precipitation and mouldiness, caused by manual operations and impacts of external factors, are avoided; the method can be applied to the biotechnology such as plant tissue culture.
Description
Technical field
The invention discloses a kind of scale preparation method of improvement White (containing mounting medium) 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 improvement White (containing mounting medium) 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: the 19 kinds of elements will improved contained by White (containing mounting medium) solid-based expand 1000 times, i.e. cupric sulfate pentahydrate 0.001g, thiamine hydrochloride 0.1g, puridoxine hydrochloride 0.1g, nicotinic acid 0.3g, potassium iodide 0.75g, boric acid 1.5g, iron sulfate 2.5g, white vitriol 2.67g, molybdic acid 3g, glycine 3g, four water manganese sulphate 6.65g, sodium dihydrogen phosphate-water 19g, potassium chloride 65g, potassium nitrate 80g, inositol 100g, sodium sulphate 200g, four water-calcium nitrate 288g, epsom salt 737g, agar powder 5000g;
2. accurate weighing: according to the result precise of 1. middle calculating, especially with the cupric sulfate pentahydrate of ten thousand/balance precise 0.001g;
3. how to mix after first few: mix according to few priority of carrying out of amount more, namely according to cupric sulfate pentahydrate, thiamine hydrochloride, puridoxine hydrochloride, nicotinic acid, potassium iodide, boric acid, iron sulfate, white vitriol, molybdic acid, glycine, four water manganese sulphates, sodium dihydrogen phosphate-water, potassium chloride, potassium nitrate, inositol, sodium sulphate, four water-calcium nitrate, epsom salt, the order of agar powder mixes, and amounts to 6509.57g;
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 6.5g 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.
A kind of scale preparation method of improvement White (containing mounting medium) 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: the 19 kinds of elements will improved contained by White (containing mounting medium) solid-based expand 1000 times, i.e. cupric sulfate pentahydrate 0.001g, thiamine hydrochloride 0.1g, puridoxine hydrochloride 0.1g, nicotinic acid 0.3g, potassium iodide 0.75g, boric acid 1.5g, iron sulfate 2.5g, white vitriol 2.67g, molybdic acid 3g, glycine 3g, four water manganese sulphate 6.65g, sodium dihydrogen phosphate-water 19g, potassium chloride 65g, potassium nitrate 80g, inositol 100g, sodium sulphate 200g, four water-calcium nitrate 288g, epsom salt 737g, agar powder 5000g;
2. accurate weighing: according to the result precise of 1. middle calculating, especially with the cupric sulfate pentahydrate of ten thousand/balance precise 0.001g;
3. how to mix after first few: mix according to few priority of carrying out of amount more, namely according to cupric sulfate pentahydrate, thiamine hydrochloride, puridoxine hydrochloride, nicotinic acid, potassium iodide, boric acid, iron sulfate, white vitriol, molybdic acid, glycine, four water manganese sulphates, sodium dihydrogen phosphate-water, potassium chloride, potassium nitrate, inositol, sodium sulphate, four water-calcium nitrate, epsom salt, the order of agar powder mixes, and amounts to 6509.57g;
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 6.5g 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 sucrose of 20g, 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 10d under 25 DEG C of illumination 12h conditions, take root good.
Claims (3)
1. improve a scale preparation method for White (containing mounting medium) solid culture matrix, it 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: the 19 kinds of elements will improved contained by White (containing mounting medium) solid-based expand 1000 times, i.e. cupric sulfate pentahydrate 0.001g, thiamine hydrochloride 0.1g, puridoxine hydrochloride 0.1g, nicotinic acid 0.3g, potassium iodide 0.75g, boric acid 1.5g, iron sulfate 2.5g, white vitriol 2.67g, molybdic acid 3g, glycine 3g, four water manganese sulphate 6.65g, sodium dihydrogen phosphate-water 19g, potassium chloride 65g, potassium nitrate 80g, inositol 100g, sodium sulphate 200g, four water-calcium nitrate 288g, epsom salt 737g, agar powder 5000g;
2. accurate weighing: according to the result precise of 1. middle calculating, especially with the cupric sulfate pentahydrate of ten thousand/balance precise 0.001g;
3. how to mix after first few: mix according to few priority of carrying out of amount more, namely according to cupric sulfate pentahydrate, thiamine hydrochloride, puridoxine hydrochloride, nicotinic acid, potassium iodide, boric acid, iron sulfate, white vitriol, molybdic acid, glycine, four water manganese sulphates, sodium dihydrogen phosphate-water, potassium chloride, potassium nitrate, inositol, sodium sulphate, four water-calcium nitrate, epsom salt, the order of agar powder mixes, and amounts to 6509.57g;
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 6.5g 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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| CN201510082346.5A CN104604713A (en) | 2015-02-16 | 2015-02-16 | Large-scale preparation method of improved White (containing fixed media) solid culture medium |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2006254832A (en) * | 2005-03-18 | 2006-09-28 | Sanei Gen Ffi Inc | Solid medium |
| CN101070531A (en) * | 2006-05-09 | 2007-11-14 | 陈曦 | Dry-powder-type culturing substrate and preparing method |
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- 2015-02-16 CN CN201510082346.5A patent/CN104604713A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2006254832A (en) * | 2005-03-18 | 2006-09-28 | Sanei Gen Ffi Inc | Solid medium |
| CN101070531A (en) * | 2006-05-09 | 2007-11-14 | 陈曦 | Dry-powder-type culturing substrate and preparing method |
Non-Patent Citations (1)
| Title |
|---|
| 仪宏等: "脱水植物组织培养基的制备", 《河北轻化工学院学报》 * |
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Application publication date: 20150513 |