CA1295569C - Method for preparing dense nutrient medium for culturing microorganisms - Google Patents
Method for preparing dense nutrient medium for culturing microorganismsInfo
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- CA1295569C CA1295569C CA000569565A CA569565A CA1295569C CA 1295569 C CA1295569 C CA 1295569C CA 000569565 A CA000569565 A CA 000569565A CA 569565 A CA569565 A CA 569565A CA 1295569 C CA1295569 C CA 1295569C
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Abstract
METHOD FOR PREPARING DENSE NUTRIENT MEDIUM
FOR CULTURING MICROORGANISMS
Abstract A method for preparing a dense nutrient medium for cul-turing microorganisms comprises copolymerization of acryl-amide, N,N'- methylenebisacrylamide and a branched polyacry-lamide with the mean molecular mass of 7.2x104 at a mass ra-tio therebetween of 7.0-15.0:0.11-0.21:0.028-0.11 respec-tively and of an initiator in a physiological solution till the formation of a gel which is then washed and impregnated with a nutrient substrate.
FOR CULTURING MICROORGANISMS
Abstract A method for preparing a dense nutrient medium for cul-turing microorganisms comprises copolymerization of acryl-amide, N,N'- methylenebisacrylamide and a branched polyacry-lamide with the mean molecular mass of 7.2x104 at a mass ra-tio therebetween of 7.0-15.0:0.11-0.21:0.028-0.11 respec-tively and of an initiator in a physiological solution till the formation of a gel which is then washed and impregnated with a nutrient substrate.
Description
12~55~9 METHOD ~R PREPARING DEN5E NUTRIENT I~EDIUM
FOR CULTURING MICROORGANISMS
The present invention relates to a method for preparing a dense nutrient medium based on a polyacrylamide gel for culturing microorganisms.
The dense nutrient medium according to the present in-vention is useful in medicine and biotechnology.
Known in the art is a method for preparing a dense nut-rient medium for culturing microorganisms by way of copoly-merization of acrylamide and N,N'-methylenebisacrylamide at a mass ratio thereof egual to 15.0-20.0:0.019-0.132 in the presence of an initiator in a physiological solution till the formation of a gel, its washing and swelling in a physio-logical solution for 16-20 hours at a temperature within the range of from 50 to 60C, followed by impregnation with a nutrient substrate.
~ his dense nutrient medium has a density approaching that of agar-agar media, it is elastic and transparent. It also ensures a good sliding of a bacteriologicalloop upon inoculation and removal of microorganisms without damaging its surface owing to a long-time operation of swelling (16--20 hours), thus substantially extending the duration of the entire process.
Known in the art is a method for preparing a nutrient medium for culturing microorganisms comprising copolymeriza-tion of acrylamide and N~N~-methylenebisacrylamide at a mass ratio thereof egual to 0.5-40.0:2.5-12.0 in the presence of an initiator in a physiological solution. The resulting poly-1~55t9 acrylamide gel is subjected to washing and impregnation witha nutrient substrate (SU, A, 977466). This dense nutrient medium has a greater sensitivity to damages during impreg-nation, sterilization and inoculation of microorganisms. The removal of individual species of microorganisms from the sur-face of such medium b~ means of a bacteriological loop is rather difficult.
In the above-discussed methods the process of copoly-merization is effected in special moulding apparatus without access of air to exclude the formation of a rough surface on the produced polyacrylamide gel hindering its use for the preparation of a nutrient medium. This process, therefore, has but a limited application in the mass-scale preparation of nutirent media, for example in Petri dishes and in other laboratory ware.
It is an obaect of the present invention to provide a method for preparing a dense nutrient medium for culturing microorganisms which would impart, to the nutrient medium, the properties similar to those of agar-agar media, while ensuring a good effect oY sliding of a bacteriological loop during inoculation of microorganisms This object is accomplished by a method f`or preparing a dense nutrient medium for culturing microorganisms which comprises copolymerization of acrylamide and N,~'-methylene-bisacrylamide in the presence of an initiator in a physio-logical solution till the formation of a gel, washing and impregnation thereof with a nutrient subs~rate, wherein, 125~55~9 according to the present invention the copolymerization of acrylamide and ~,N'-methylenebisacrylamide is carried out in the presence of a branched polyacrylamide with a mean mole-cular mass o~ 7,2x104 at a mass ratio thereof equal to 7.0-15.0:0.11-0,21:0.028-0.11 respectively.
~ he method according to the present invention provides for a good quality of the nutrient medium at a simplified process of its manufacture. ~he process of copolymerization, owing to the use of the structurization agent, is readily implementable in the air atmosphere without the necessity of using special equipment, ~ o acceletate the reaction of copolymerization, it is advisable to use an agueous solution of the above-mentioned branched polyacrylamide. It is desirable to use, for the imp-regnation of the polyacrylamide gel, a nutrient substrate containing polyethylene oxide in an amount of 0.02 to 0,2 g/l.
~his contributes to the formatioD of such a superficial mono-layer on the surface of the medium which ensures a clearly pronounced effect of sliding of a bacteriological loop in inoculation of microorganisms.
~ he dense nutrient medium for culturing microorganisms according to the present invention is prepared in the follow-ing manner.
A reaction mixture is prepared which contains solutions of the starting monomers, a branched polyacrylamide with the molecular mass of 7.2x104 and initiators in a physiolo-gical solution. It is also possible to ùse an aqueous solu-1~55~
tion of the above-specified branched polyacrylamide. The ra-tio of acrylamide and the branched polyacrylamide in the reac-tion mixture is 7.0-1.5:0.028-0.11 respectively.
As the physiological solution use is made of a 0.5% or a 0.9% solution of sodium chloride, a 5% agueous solution of glucose, the Ringer-Lock solution, Hanks solution, Earl so-lution and the like. ~he copolymerization is conducted in the air in a Petri dish~r ~ thaother vessels made from diffe-rent materials.
The process of copolymerization in the presence of a branched polyacrylamide employed in the above-specified pro-portions enables the production of a polyacrylamide gel with a density close to that of the agar base. Furthermore, the use of a branched polyacrylamide results in an additional structurization of the polyacrylamide ~el, thus enabling the formation of an even smooth surface thereof upon polymeriza-tion in Petri dishes in the air. ~reviously such an effect could be obtained only upon polymerization in special appa-ratus without access of air and with the use of inert gases.
In the method for preparing nutrient media according to the present invention the stage of the gel sw ~ling for 16-20 hours is excluded.
All the above-mentioned advantages substantially simpli-fy the process for the preparation of a nutrient medium.
After washing, the polyacrylamide gel is subjected to sterilization by way of a thermal, radiation or chemical treatment. The impregnation of the above-mentioned gel with lZ95S69 substrates for nutrition of microorganisms can be effected both before and after sterilization. It is advisable to use nutrient substrates containing polyethylene oxide with a mo-lecular mass of from 500,000 onwards in an amount of from 0.02 to 0.2.
The substrate composition is defined by the needs of particular groups or species of microorganisms and cells.
~ or the determination of the quality of nutrient media, as well as for the study of biological properties of micro-organisms, generally accepted investigation procedures are employed.
Specific examples illustrating some particular embodi-ments of the method according to the present invention are given hereinbelow.
~ ample 1 For the production of a ~el five solutions A,B, C, D
and ~ are prepared following the procedure specified herein-below (the component amounts are given per 100 ml of the so-lution).
1. Preparation of solution A
0.5 ml of N,N,N',N'-tetramethylethylenediamine (TEMED) is dissolved in 99.5 ml of a physiological solution.
FOR CULTURING MICROORGANISMS
The present invention relates to a method for preparing a dense nutrient medium based on a polyacrylamide gel for culturing microorganisms.
The dense nutrient medium according to the present in-vention is useful in medicine and biotechnology.
Known in the art is a method for preparing a dense nut-rient medium for culturing microorganisms by way of copoly-merization of acrylamide and N,N'-methylenebisacrylamide at a mass ratio thereof egual to 15.0-20.0:0.019-0.132 in the presence of an initiator in a physiological solution till the formation of a gel, its washing and swelling in a physio-logical solution for 16-20 hours at a temperature within the range of from 50 to 60C, followed by impregnation with a nutrient substrate.
~ his dense nutrient medium has a density approaching that of agar-agar media, it is elastic and transparent. It also ensures a good sliding of a bacteriologicalloop upon inoculation and removal of microorganisms without damaging its surface owing to a long-time operation of swelling (16--20 hours), thus substantially extending the duration of the entire process.
Known in the art is a method for preparing a nutrient medium for culturing microorganisms comprising copolymeriza-tion of acrylamide and N~N~-methylenebisacrylamide at a mass ratio thereof egual to 0.5-40.0:2.5-12.0 in the presence of an initiator in a physiological solution. The resulting poly-1~55t9 acrylamide gel is subjected to washing and impregnation witha nutrient substrate (SU, A, 977466). This dense nutrient medium has a greater sensitivity to damages during impreg-nation, sterilization and inoculation of microorganisms. The removal of individual species of microorganisms from the sur-face of such medium b~ means of a bacteriological loop is rather difficult.
In the above-discussed methods the process of copoly-merization is effected in special moulding apparatus without access of air to exclude the formation of a rough surface on the produced polyacrylamide gel hindering its use for the preparation of a nutrient medium. This process, therefore, has but a limited application in the mass-scale preparation of nutirent media, for example in Petri dishes and in other laboratory ware.
It is an obaect of the present invention to provide a method for preparing a dense nutrient medium for culturing microorganisms which would impart, to the nutrient medium, the properties similar to those of agar-agar media, while ensuring a good effect oY sliding of a bacteriological loop during inoculation of microorganisms This object is accomplished by a method f`or preparing a dense nutrient medium for culturing microorganisms which comprises copolymerization of acrylamide and N,~'-methylene-bisacrylamide in the presence of an initiator in a physio-logical solution till the formation of a gel, washing and impregnation thereof with a nutrient subs~rate, wherein, 125~55~9 according to the present invention the copolymerization of acrylamide and ~,N'-methylenebisacrylamide is carried out in the presence of a branched polyacrylamide with a mean mole-cular mass o~ 7,2x104 at a mass ratio thereof equal to 7.0-15.0:0.11-0,21:0.028-0.11 respectively.
~ he method according to the present invention provides for a good quality of the nutrient medium at a simplified process of its manufacture. ~he process of copolymerization, owing to the use of the structurization agent, is readily implementable in the air atmosphere without the necessity of using special equipment, ~ o acceletate the reaction of copolymerization, it is advisable to use an agueous solution of the above-mentioned branched polyacrylamide. It is desirable to use, for the imp-regnation of the polyacrylamide gel, a nutrient substrate containing polyethylene oxide in an amount of 0.02 to 0,2 g/l.
~his contributes to the formatioD of such a superficial mono-layer on the surface of the medium which ensures a clearly pronounced effect of sliding of a bacteriological loop in inoculation of microorganisms.
~ he dense nutrient medium for culturing microorganisms according to the present invention is prepared in the follow-ing manner.
A reaction mixture is prepared which contains solutions of the starting monomers, a branched polyacrylamide with the molecular mass of 7.2x104 and initiators in a physiolo-gical solution. It is also possible to ùse an aqueous solu-1~55~
tion of the above-specified branched polyacrylamide. The ra-tio of acrylamide and the branched polyacrylamide in the reac-tion mixture is 7.0-1.5:0.028-0.11 respectively.
As the physiological solution use is made of a 0.5% or a 0.9% solution of sodium chloride, a 5% agueous solution of glucose, the Ringer-Lock solution, Hanks solution, Earl so-lution and the like. ~he copolymerization is conducted in the air in a Petri dish~r ~ thaother vessels made from diffe-rent materials.
The process of copolymerization in the presence of a branched polyacrylamide employed in the above-specified pro-portions enables the production of a polyacrylamide gel with a density close to that of the agar base. Furthermore, the use of a branched polyacrylamide results in an additional structurization of the polyacrylamide ~el, thus enabling the formation of an even smooth surface thereof upon polymeriza-tion in Petri dishes in the air. ~reviously such an effect could be obtained only upon polymerization in special appa-ratus without access of air and with the use of inert gases.
In the method for preparing nutrient media according to the present invention the stage of the gel sw ~ling for 16-20 hours is excluded.
All the above-mentioned advantages substantially simpli-fy the process for the preparation of a nutrient medium.
After washing, the polyacrylamide gel is subjected to sterilization by way of a thermal, radiation or chemical treatment. The impregnation of the above-mentioned gel with lZ95S69 substrates for nutrition of microorganisms can be effected both before and after sterilization. It is advisable to use nutrient substrates containing polyethylene oxide with a mo-lecular mass of from 500,000 onwards in an amount of from 0.02 to 0.2.
The substrate composition is defined by the needs of particular groups or species of microorganisms and cells.
~ or the determination of the quality of nutrient media, as well as for the study of biological properties of micro-organisms, generally accepted investigation procedures are employed.
Specific examples illustrating some particular embodi-ments of the method according to the present invention are given hereinbelow.
~ ample 1 For the production of a ~el five solutions A,B, C, D
and ~ are prepared following the procedure specified herein-below (the component amounts are given per 100 ml of the so-lution).
1. Preparation of solution A
0.5 ml of N,N,N',N'-tetramethylethylenediamine (TEMED) is dissolved in 99.5 ml of a physiological solution.
2. Preparation of solution B
0.735 g of N,N'-methylenebisacrylamide (MBA) is dissolv-ed in 50 ml of a physiological solution, heated to the tem-perature of 60C, added with 24.5 g of acrylamide (AA) and stirred till a complete dissolution. ~he resulting solution is filtered and brought to the volume of 100 ml by means of a physiological solution.
125'55~;~
0.735 g of N,N'-methylenebisacrylamide (MBA) is dissolv-ed in 50 ml of a physiological solution, heated to the tem-perature of 60C, added with 24.5 g of acrylamide (AA) and stirred till a complete dissolution. ~he resulting solution is filtered and brought to the volume of 100 ml by means of a physiological solution.
125'55~;~
3. Preparation of solution C
0.2 g of ammonium persulphate (AP) is dissolved in 50 ml of a physiological solution and brought to the volume of 100 ml.
0.2 g of ammonium persulphate (AP) is dissolved in 50 ml of a physiological solution and brought to the volume of 100 ml.
4. Preparation of solution D
10.0g of a branched polyacrylamide (BPA) with the mole-cular mass of 7.2x104 are dissolved in 50 ml of a physiolo-gical solution at a temperature of 50-60C. The volume of the solution is brought to 1GOml.
10.0g of a branched polyacrylamide (BPA) with the mole-cular mass of 7.2x104 are dissolved in 50 ml of a physiolo-gical solution at a temperature of 50-60C. The volume of the solution is brought to 1GOml.
5. Preparation of solution E
0.5 ml of solution D is mixed with ~9.5 ml of solu-tion C and thoroughly intermixed.
The reaction mixture is prepared from the solutions emp-loyed in the followingr ratio: A:B:~ = 1:2:4 respectively which corresponds to the mass ratio of acrylamide to N,N'-methylenebisacrylamide and to the branched polyacrylamide equal to 7.0:0.21:0.028. The mixture is poured into Petri dishes ana allowed to stay in the air. ~he copolymeriza-tion occurs for 10-15 minutes till the formation of a gel which is then washed with a physiological solution for one hour.
A dense nutrient medium is prepared by impregnation of the resulting ~el with the Hottingen tryptic digestion broth.
The nutrient medium is inoculated with Staphylococcus aureus and incubated at the temperature of 37C for 24 hours. The dense nutrient medium has good adhesion pro-lZ955~,9 perties, density, transparencey and elasticity, it has asmooth surface ensuring the effect of sliding of a bacterio-logical loop upon inoculation of the microorganisms without damaging the surface. The grown microorganisms demonstrate characteristic cultural ard morphological features.
Example 2 For the preparation of a polyacrylamide gel solutions A, B, C and D are used likewise in the foregoing Example 1.
The reaction mixture is prepared from the solutions at the ratios of A:B:E = 1:2:4 which corresponds to the mass ratio of acrylamide, N,N'-methylenebisacrylamide and the branched polyacrylamide of 7.0:0.21:0.057. The mixture is poured into Petri dishes and allowed for polymerization in the air. The copolymerization occurs for 5-10 minutes un-til a gel is formed which is then washed with a physiologi-cal solution for one hour.
The resulting gel is impregnated with a meat-peptone broth for 3 hours at the temperature of 56C, followed by sterilization for 30 minutes.
The nutrient medium is inoculated with colibacillus and incubated at the temperature of 37C for 24 hours. The grown microorganisms reveal cultural and morphological fea-tures characteristic therefor. The dense nutrient medium has good adhesion properties, density, transparency, elasti-city; it has a smooth surface ensuring the effect of sliding for a bacteriological loop upon inoculation of the microor-ganisms without damaging the surface.
12955~9 Example 3 For the pleparation of a polyacrylamide gel three solu-tions A, B and C are obtained according to the following pro-cedure.
1. Preparation of solution A
0.23 ml of N,N,N',N'-tetramethylethylenediamine is dis-solved in 50 ml of a physiological solution and brought to the volume of 100 ml.
2. Preparation of solution B
0.4 g of N,N'-methylenebisacrylamide is dissolved in 50 ml of a physiological solution, heated to the temperature of 60C, added with 53 g of acrylamide and stirred till a complete dissolution. The resulting solution is filtered, brought to the volume of 100 ml.
3. Preparation of solutions C and D
The solutions C and D are prepared in a manner similar to that described in Example 1 hereinbefore.
4. Preparation of solution E
2 ml of solution D are introduced into 98 ml of solu-tion C and thoroughly intermixed.
A reaction mixture is produced from the resulting pre-pared~solutions. To this end, the solutions are mixed in the ratio of A:B:E = 1:2:4 which corresponds to the mass ratio of acrylamide, N,N'-methyleneacrylamide and the branched polyacrylamide of 15.0:0.11:0.11 and poured into Petri dishes. The copolymerization is conducted in the air for 10-15 minutes till a gel is formed. The resulting gel is _ 9 _ washed with physiological solution for one hour. ~he gel is then subjected to impregnation with a meat-peptone broth for 3 hours at a temperature of 50-60C and a spore culture is inoculated after sterilization.
~ he resultinr dense nutrient medium has ~ood adhesion properties, elasticity, density, transparency; it also fea-tures a smooth s~ face, a clearly pronounced effect of slid-ing of a bacteriolo~ical loop upon inoculation of microorga-nisms. The grown culture is typical.
Example 4 A polyacrylamide gel is produced in a manner similar to that described in Example 1 hereinbefore, while the impreg-nation is effected in the Hottinger tryptic d~stion broth containing 0.2 g/l of polyethyleneoxide for 3 hours at the temperature of 56C. The impLegDated gel is sterilized for 30 minutes and inoculated with Staphylococcus aureus.
The resulting dense nutrient medium exhibits good adhe-sion properties, elasticity, density, transparency and a clearly pronounced effect of JlidiL-~ upon inoculation of the microorganisms by means of a bacteriological loop without damaging the culture surface. The grown culture is typical.
Example 5 A polyacrylamide gel is produced as in Example 3 herein-before, while the impregnation is effected by means of a ~et-peptone broth containing 0.02 g/l of polyethyleneoxide for 3 hours at the temperature of 56C, followed by sterili-zation for 30 minutes lZ95Stj9 - 10 _ ~ he resulting dense nutrient medium i`eatures good adhe-sion properties, ela~ticity, density, transparency; it also has a clearly pronounced sliding ef~ect upon inoculation of the microorganisms by means of a bacteriological loop with-out damaging the culture surface thereby.
As the test-microbe colibacillus is used.
The culture gro~s in the form of typical colonies.
0.5 ml of solution D is mixed with ~9.5 ml of solu-tion C and thoroughly intermixed.
The reaction mixture is prepared from the solutions emp-loyed in the followingr ratio: A:B:~ = 1:2:4 respectively which corresponds to the mass ratio of acrylamide to N,N'-methylenebisacrylamide and to the branched polyacrylamide equal to 7.0:0.21:0.028. The mixture is poured into Petri dishes ana allowed to stay in the air. ~he copolymeriza-tion occurs for 10-15 minutes till the formation of a gel which is then washed with a physiological solution for one hour.
A dense nutrient medium is prepared by impregnation of the resulting ~el with the Hottingen tryptic digestion broth.
The nutrient medium is inoculated with Staphylococcus aureus and incubated at the temperature of 37C for 24 hours. The dense nutrient medium has good adhesion pro-lZ955~,9 perties, density, transparencey and elasticity, it has asmooth surface ensuring the effect of sliding of a bacterio-logical loop upon inoculation of the microorganisms without damaging the surface. The grown microorganisms demonstrate characteristic cultural ard morphological features.
Example 2 For the preparation of a polyacrylamide gel solutions A, B, C and D are used likewise in the foregoing Example 1.
The reaction mixture is prepared from the solutions at the ratios of A:B:E = 1:2:4 which corresponds to the mass ratio of acrylamide, N,N'-methylenebisacrylamide and the branched polyacrylamide of 7.0:0.21:0.057. The mixture is poured into Petri dishes and allowed for polymerization in the air. The copolymerization occurs for 5-10 minutes un-til a gel is formed which is then washed with a physiologi-cal solution for one hour.
The resulting gel is impregnated with a meat-peptone broth for 3 hours at the temperature of 56C, followed by sterilization for 30 minutes.
The nutrient medium is inoculated with colibacillus and incubated at the temperature of 37C for 24 hours. The grown microorganisms reveal cultural and morphological fea-tures characteristic therefor. The dense nutrient medium has good adhesion properties, density, transparency, elasti-city; it has a smooth surface ensuring the effect of sliding for a bacteriological loop upon inoculation of the microor-ganisms without damaging the surface.
12955~9 Example 3 For the pleparation of a polyacrylamide gel three solu-tions A, B and C are obtained according to the following pro-cedure.
1. Preparation of solution A
0.23 ml of N,N,N',N'-tetramethylethylenediamine is dis-solved in 50 ml of a physiological solution and brought to the volume of 100 ml.
2. Preparation of solution B
0.4 g of N,N'-methylenebisacrylamide is dissolved in 50 ml of a physiological solution, heated to the temperature of 60C, added with 53 g of acrylamide and stirred till a complete dissolution. The resulting solution is filtered, brought to the volume of 100 ml.
3. Preparation of solutions C and D
The solutions C and D are prepared in a manner similar to that described in Example 1 hereinbefore.
4. Preparation of solution E
2 ml of solution D are introduced into 98 ml of solu-tion C and thoroughly intermixed.
A reaction mixture is produced from the resulting pre-pared~solutions. To this end, the solutions are mixed in the ratio of A:B:E = 1:2:4 which corresponds to the mass ratio of acrylamide, N,N'-methyleneacrylamide and the branched polyacrylamide of 15.0:0.11:0.11 and poured into Petri dishes. The copolymerization is conducted in the air for 10-15 minutes till a gel is formed. The resulting gel is _ 9 _ washed with physiological solution for one hour. ~he gel is then subjected to impregnation with a meat-peptone broth for 3 hours at a temperature of 50-60C and a spore culture is inoculated after sterilization.
~ he resultinr dense nutrient medium has ~ood adhesion properties, elasticity, density, transparency; it also fea-tures a smooth s~ face, a clearly pronounced effect of slid-ing of a bacteriolo~ical loop upon inoculation of microorga-nisms. The grown culture is typical.
Example 4 A polyacrylamide gel is produced in a manner similar to that described in Example 1 hereinbefore, while the impreg-nation is effected in the Hottinger tryptic d~stion broth containing 0.2 g/l of polyethyleneoxide for 3 hours at the temperature of 56C. The impLegDated gel is sterilized for 30 minutes and inoculated with Staphylococcus aureus.
The resulting dense nutrient medium exhibits good adhe-sion properties, elasticity, density, transparency and a clearly pronounced effect of JlidiL-~ upon inoculation of the microorganisms by means of a bacteriological loop without damaging the culture surface. The grown culture is typical.
Example 5 A polyacrylamide gel is produced as in Example 3 herein-before, while the impregnation is effected by means of a ~et-peptone broth containing 0.02 g/l of polyethyleneoxide for 3 hours at the temperature of 56C, followed by sterili-zation for 30 minutes lZ95Stj9 - 10 _ ~ he resulting dense nutrient medium i`eatures good adhe-sion properties, ela~ticity, density, transparency; it also has a clearly pronounced sliding ef~ect upon inoculation of the microorganisms by means of a bacteriological loop with-out damaging the culture surface thereby.
As the test-microbe colibacillus is used.
The culture gro~s in the form of typical colonies.
Claims (3)
1. A method for preparing a dense nutrient medium for culturing microorganisms comprising copolymerization of ac-rylamide and N,N'-methylenebisacrylamide in the presence of a branched polyacrylamide with the mean molecular mass of 7.2x104 at a mass ratio therebetween of 7.0-15.0:
:0.11-0.21:0.028-0.11 respectively and of an initiator in a physiological solution till the formation of a gel, washing and impregnation thereof with a nutrient substrate.
:0.11-0.21:0.028-0.11 respectively and of an initiator in a physiological solution till the formation of a gel, washing and impregnation thereof with a nutrient substrate.
2. A method according to Claim 1, wherein an aqueous solution of said polyacrylamide is used.
3. A method according to Claim 1, wherein the impregna-tion of said gel is effected by means of a nutrient substrate containing polyethyleneoxide in an amount of from 0.02 to 0.2 g/l.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| SU874278310A RU1837059C (en) | 1987-06-25 | 1987-06-25 | Method for producing solid nutrient medium for cultivation of microorganism |
| SU4278310 | 1987-06-25 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1295569C true CA1295569C (en) | 1992-02-11 |
Family
ID=21317183
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000569565A Expired - Fee Related CA1295569C (en) | 1987-06-25 | 1988-06-15 | Method for preparing dense nutrient medium for culturing microorganisms |
Country Status (5)
| Country | Link |
|---|---|
| CN (1) | CN1030428A (en) |
| CA (1) | CA1295569C (en) |
| ES (1) | ES2007242A6 (en) |
| RU (1) | RU1837059C (en) |
| YU (1) | YU121588A (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| BR112015012391A2 (en) * | 2012-12-19 | 2017-07-11 | Nestec Sa | Dairy compound dessert and its preparation process |
-
1987
- 1987-06-25 RU SU874278310A patent/RU1837059C/en active
-
1988
- 1988-06-15 CA CA000569565A patent/CA1295569C/en not_active Expired - Fee Related
- 1988-06-23 YU YU121588A patent/YU121588A/en unknown
- 1988-06-23 CN CN 88106010 patent/CN1030428A/en active Pending
- 1988-06-24 ES ES8801978A patent/ES2007242A6/en not_active Expired
Also Published As
| Publication number | Publication date |
|---|---|
| YU121588A (en) | 1989-06-30 |
| CN1030428A (en) | 1989-01-18 |
| RU1837059C (en) | 1993-08-30 |
| ES2007242A6 (en) | 1989-06-01 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| MKLA | Lapsed |