CN104604719A

CN104604719A - Method for preparing WPM (carbon-source-containing) solid culture medium in large scale

Info

Publication number: CN104604719A
Application number: CN201510082583.1A
Authority: CN
Inventors: 钱夕刚
Original assignee: Individual
Current assignee: Individual
Priority date: 2015-02-16
Filing date: 2015-02-16
Publication date: 2015-05-13

Abstract

The invention discloses a method for preparing a WPM (carbon-source-containing) solid culture medium in a large scale and belongs to the technical field of plant biology. The method for preparing the WPM (carbon-source-containing) solid culture medium in the large scale comprises the following steps: firstly calculating demanded quantities of eighteen components contained in the WPM (carbon-source-containing) solid culture medium after being expanded by 1000 times; accurately weighing expanded quantities of medicaments, wherein sodium molybdate dihydrate and copper sulphate pentahydrate are accurately weighed by utilizing a ten-thousandth balance; successively mixing the eighteen components according to gradually increasing demanded quantities, namely the two components with the lowest demanded quantities are mixed firstly, then the component with the next-to-the-lowest demanded quantity is added, uniform mixing is carried out, and the process is repeated; sealing every 250g of solid culture medium in one fixed plastic container; and opening the plastic containers respectively as required during application. The method for preparing the WPM (carbon-source-containing) solid culture medium in the large scale has the advantages that the WPM (carbon-source-containing) solid culture medium expanded by 1000 times can be prepared, the characteristics of easiness and speediness in operation, simplicity in application, small error in different batches and small intra-batch error can be realized, and the phenomena, such as big error, precipitation and moulding, which are caused by manual operations such as calculating and weighing and external factors such as weather, temperature and moisture during preparation of WPM liquid culture medium can be avoided, so that the method for preparing the WPM (carbon-source-containing) solid culture medium in the large scale can be applied to biotechnical uses such as plant tissue culture.

Description

A kind of scale preparation method of WPM (carbonaceous sources) solid culture matrix

Technical field

The invention discloses the scale preparation method of a kind of WPM (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 WPM (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: 18 kinds of elements contained by WPM (carbonaceous sources) solid culture medium are expanded 1000 times, i.e. cupric sulfate pentahydrate 0.25g, Sodium Molybdate Dihydrate 0.25g, nicotinic acid 0.5g, puridoxine hydrochloride 0.5g, thiamine hydrochloride 1g, glycine 2g, boric acid 6.2g, white vitriol 8.6g, four water manganese sulphate 22.5g, iron edetate 36.5g, calcium chloride dihydrate 96g, inositol 100g, potassium dihydrogen phosphate 170g, magnesium sulfate 370g, ammonium nitrate 400g, four water-calcium nitrate 556g, potassium sulfate 990g, soft white sugar 30000g;

2. accurate weighing: according to the result precise of 1. middle calculating;

3. how to mix after first few: mix according to few priority of carrying out of amount more, namely according to cupric sulfate pentahydrate, Sodium Molybdate Dihydrate, nicotinic acid, puridoxine hydrochloride, thiamine hydrochloride, glycine, boric acid, white vitriol, four water manganese sulphates, iron edetate, calcium chloride dihydrate, inositol, potassium dihydrogen phosphate, magnesium sulfate, ammonium nitrate, four water-calcium nitrate, potassium sulfate, the order of soft white sugar mixes, and amounts to 32760.3g;

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 32.76g 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 WPM (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:

2. accurate weighing: according to the result precise of 1. middle calculating;

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.Photinia glabra stem section is inoculated on superclean bench, after cultivating 45d under 25 DEG C of 12h illumination conditions, stem section well-grown and occur a large amount of axillalry bud.

Claims

1. a scale preparation method for WPM (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:

2. accurate weighing: according to the result precise of 1. middle calculating;

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.