CN104542310A

CN104542310A - Scale preparation method of 1/2MS (containing fixed medium) basic solid culture substrate

Info

Publication number: CN104542310A
Application number: CN201510082628.5A
Authority: CN
Inventors: 钱夕刚
Original assignee: Individual
Current assignee: Individual
Priority date: 2015-02-16
Filing date: 2015-02-16
Publication date: 2015-04-29

Abstract

The invention discloses a scale preparation method of a 1/2MS (containing a fixed medium) basic solid culture substrate, belonging to the technical field of plant biology. The scale preparation method disclosed by the invention comprises the following steps of: calculating the demanded quantity after 19 elements contained in the 1/2MS (containing the fixed medium) basic solid culture substrate are expanded by 1000 times at first; accurately weighing the enlarged drug amount, wherein CuSO4.5H2O and CoCl12.6H2O need to be accurately weighed by using a one over ten-thousand balance; sequentially mixing the 19 elements according to the demanded quantity, namely, mixing two least elements at first, and then, uniformly mixing by adding second little elements, by parity of reasoning; sealing every 250 g of the solid culture substrate in a fixed plastic container; and respectively unsealing and using according to requirements. By means of the scale preparation method disclosed by the invention, the 1/2MS (containing the fixed medium) basic solid culture substrate expanded by 1000 times can be prepared; the scale preparation method has the characteristics of being convenient and rapid to operate, simple to apply, different in batch and low in error in batch; the phenomena of high error, precipitation, mildew and the like due to manual operation, such as calculating and weighing, and external factors, such as weather, temperature and water content, in the event of preparing a 1/2MS liquid culture medium, can be avoided; and the scale preparation method disclosed by the invention can be used in biotechnologies, such as plant tissue culture and the like.

Description

The scale preparation method of the basic solid culture matrix of a kind of 1/2MS (containing mounting medium)

Technical field

The invention discloses the scale preparation method of the basic solid culture matrix of a kind of 1/2MS (containing mounting medium), 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

The scale preparation method of the basic solid culture matrix of a kind of 1/2MS (containing mounting medium), 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: element in 19 contained by basic for 1/2MS (containing mounting medium) solid-based is expanded 1000 times, i.e. CoCL2 6H2O and each 0.025g of cupric sulfate pentahydrate, thiamine hydrochloride 0.1g, Sodium Molybdate Dihydrate 0.25g, 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 85g, epsom salt 195g, calcium chloride dihydrate 220g, ammonium nitrate 825g, potassium nitrate 950g, agar powder 5000g,

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, thiamine hydrochloride, Sodium Molybdate Dihydrate, 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 agar powder mixes, and amounts to 7442.83g;

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 7.44g 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 the basic solid culture matrix of a kind of 1/2MS (containing mounting medium) 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:

1L solution after above-mentioned substrate preparation is added the sucrose of 30g, 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 15d under 25 DEG C of illumination 12h conditions, take root good.

Claims

1. a scale preparation method for the basic solid culture matrix of 1/2MS (containing mounting medium), 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:

Accurate calculating: element in 19 contained by basic for 1/2MS (containing mounting medium) solid-based is expanded 1000 times, i.e. CoCL2 6H2O and each 0.025g of cupric sulfate pentahydrate, thiamine hydrochloride 0.1g, Sodium Molybdate Dihydrate 0.25g, 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 85g, epsom salt 195g, calcium chloride dihydrate 220g, ammonium nitrate 825g, potassium nitrate 950g, agar powder 5000g;

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;

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, thiamine hydrochloride, Sodium Molybdate Dihydrate, 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 agar powder mixes, and amounts to 7442.83g;

Sealing: the requirement of mixed solid matrix according to 250g every bottle is sub-packed in plastic containers, and seals;

Unpacking uses: as required, add the above-mentioned matrix 7.44g of precise in every 1L distilled water, and 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.