CN104604700A

CN104604700A - Large-scale production method for positive 14 (containing solid medium) solid culture medium

Info

Publication number: CN104604700A
Application number: CN201510082186.4A
Authority: CN
Inventors: 钱家静
Original assignee: Individual
Current assignee: Individual
Priority date: 2015-02-16
Filing date: 2015-02-16
Publication date: 2015-05-13

Abstract

The invention discloses a large-scale production method for positive 14 (containing a solid medium) solid culture medium, belonging to the field of plant biotechnologies. The large-scale production method comprises the following steps: firstly calculating the demanded quantities of 18 elements contained in the positive 14 (containing the solid medium) solid culture medium after 1000 times of amplification; accurately weighing amount of the amplified medicine, wherein CuSO4.5H2O and CoCl2.6H2O need to be accurately weighed by a one over ten-thousand balance; sequentially mixing the 18 elements according to the demanded quantities, namely, mixing the two fewest elements at first, and then adding the second fewest elements and uniformly mixing, and the like; sealing per 250g of the solid culture medium in a solid plastic container; and unsealing for use as needed respectively. The positive 14 (containing the solid medium) solid culture medium after 1000 times of amplification can be prepared through the method, has the characteristics of being convenient to operate, rapid, simple to apply, and having small errors of different batches and in the batches, avoids the phenomena of large errors, precipitation, mildewing and the like caused by the influences of manual operations of calculation, weighing and the like, and external factors such as weather, temperature, water content and the like during preparation for a positive 14 liquid culture medium, as well as can be applied to biotechnological applications of plant tissue culture and the like.

Description

A kind of scale preparation method of positive 14 (containing mounting medium) solid culture matrix

Technical field

The invention discloses a kind of scale preparation method of positive 14 (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 positive 14 (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: 18 kinds of elements contained by positive 14 (containing mounting medium) solid culture medium are expanded 1000 times, i.e. CoCL2 6H2O and each 0.025g of cupric sulfate pentahydrate, Sodium Molybdate Dihydrate 0.25g, potassium iodide 0.75g, nicotinic acid 1g, puridoxine hydrochloride 1g, glycine 2g, white vitriol 2g, boric acid 3g, thiamine hydrochloride 10g, manganese sulfate monohydrate 10g, iron edetate 36.5g, calcium chloride dihydrate 150g, ammonium sulfate 150g, epsom salt 450g, potassium dihydrogen phosphate 600g, potassium nitrate 3000g, 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, Sodium Molybdate Dihydrate, potassium iodide, nicotinic acid, puridoxine hydrochloride, glycine, white vitriol, boric acid, thiamine hydrochloride, manganese sulfate monohydrate, iron edetate, calcium chloride dihydrate, ammonium sulfate, epsom salt, potassium dihydrogen phosphate, potassium nitrate, the order of agar powder mixes, and amounts to 9416.55g;

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

5. to break seal use: as required, add the above-mentioned matrix 9.42g 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 positive 14 (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:

1L solution after above-mentioned substrate preparation is added the sucrose of 15g, 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 18d under 25 DEG C of illumination 12h conditions, root is many and take root good.

Claims

1. a scale preparation method for positive 14 (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.

2. method according to claim 1, is characterized in that, described five steps concrete operations are as follows:

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.