preparation and sterilization of culture media ppt

1) Standard mode: Heating up, sterilization and cooling down to dispensing temperature. Preparation of sterile media, containers and small instruments. The filtered water is now added for appropriate dilution of the media. 5.2.7 Sterilize the media by autoclaving at 121°C for 15 minutes at 15 lbs pressure unless otherwise specified. Sterilization during autoclaving depends mainly on temperature. A number 3 Preparation of sterile media, containers and small instruments. Steam enters into a … 8. Contents. �:�s�M��:�y��Yt� 9 Extraction of secondary metabolites from callus Preparation and sterilization of culture media are very important to prevent contamination of the unwanted microorganisms. Autoclave sterilization for 15 minutes at 15 pounds of pressure and at 121 °C is The most popular method of sterilizing culture media and apparatus is by autoclaving the material using steam/dry sterilization. Media sterilization is carried out with the autoclave, basically a huge steam cooker. Culture media should contain at least 25-60 mM of inorganic nitrogen for satisfactory plant cell growth. endstream endobj startxref %PDF-1.5 %���� h��X�o�8�W�q�г����@��[��KoP��x�qy�vq�?R��M�]n�B@H�H����P���P�AC 7�0B���) A���J"3ey*��j���I�����0�4t��0�a�kv3NMq:0�)r���`LK���&�(I��h4��bJ�B? Control of culture media, in terms of appropriate records through to plate reading, forms an important part of data integrity in the microbiology laboratory (as assessed by Saha (2016) and Sandle (2016) (2, 3). 4Dx�.o���A��7Qo����Lz S��v�iF�@�����*��� ` � Sterilization of Culture Media Nutrient media as initially prepared contain a variety of different vegetative cells and spores, derived from the constituents of the culture me-dium, the water, and the vessel. Containers used for media must have vented tops and should be capable of holding at least 20% more than the intended volume of medium, to allow for expansion during sterilization. 0 endstream endobj 156 0 obj <>/Metadata 9 0 R/PageLayout/OneColumn/Pages 153 0 R/StructTreeRoot 18 0 R/Type/Catalog>> endobj 157 0 obj <>/ExtGState<>/Font<>/XObject<>>>/Rotate 0/StructParents 0/Tabs/S/Type/Page>> endobj 158 0 obj <>stream Sterilization methods include; autoclaving, dry-heat, filtration, UV exposure and ethylene oxide. Preparation and sterilization of culture media is important to prevent contamination of the unwanted microorganisms inside the media. h�bbd``b`VӀ�:�`�l .�eQ $XSAD+��D����� �= cd�2��+����� The separation of the cooling water system and the sterilization chamber makes it unlikely that culture media gets contaminated by cooling water. 9. … The Sterilization Service of the CIB is devoted to sterilize, through dry or wet heat, the working material and wastes of … Introduction 151. The chemical methods (by using disinfectants) and radiation procedures (by using UV rays, y rays, X-rays) are not commonly used for media sterilization. Microbes can use the nutrients of culture media as their food is necessary for cultivating them in vitro. As the tissue culture is being carried out in highly aseptic condition, all the usable, media and solutions are required to be sterilized thoroughly. Preparation of Culture Media 377. Following sterilization in a flask or bottle, the media is poured into plates using aseptic technique, preferably in a sterile cabinet (laminar flow hood). Add heat labile constituents after autoclaving. %PDF-1.5 %���� 2) Chocolate agar mode: After the first sterilization phase, blood is added and the medium heated up again. Although sterilization of culture media is best carried out in a steam autoclave at temperature around 121C it has recognized that damage is caused to the medium by the heating process. Enriched media are the artificial culture media that are enriched with whole blood, lyze blood, serum, extra peptones, special extracts or vitamins to support the growth of pathogens which require additional nutrients or growth stimulants e.g. HEAD OF STERILIZATION AND CULTURE MEDIA PREPARATION FACILITY: Eduardo Díaz Fernández. Autoclaving Culture vessels, etc. Dispense the medium into the culture vessels before (or after) autoclaving according to your application. Plant tissue culture media are generally sterilized by autoclaving at 121 °C and 1.05 kg/cm 2 (15-20 psi). 3 Preparation of media ms medium. Maintenance of aseptic conditions. Sterilize the medium in a validated autoclave at 1 kg/cm2 (15 psi), 121 °C, for the time period described under Sterilization of Media Protocol. The media must be free from contamination before use in fermentation. 4 Surfacesterilization of explants. %%EOF 5 Preparation of explant for callusinduction. STERILIZATION AND CULTURE MEDIA PREPARATION FACILITY. Table of contents Preparation of culture media, agar plates, antibiotics and general necessities 1 Preparation of Culture Media 3 1.1 Luria-Bertani (LB) Medium 3 1.1.1 Materials 3 1.1.2 Setup & Protocol 3 1.2 2YT 3 1.2.1 Materials 3 1.2.2 Setup & Protocol 4 2 Agar Plates 4 2.1 Preparation of LB-agar 4 2.1.1 Materials 4 2.1.2 Setup & Protocol 4 2.2 Pouring the plates 4 520 0 obj <> endobj The minimum times required for sterilization of different volumes of medium are listed below. 198 0 obj <>stream Besides, different types of agar are needed for the cultivation of different types of microorganisms. 1 Sterilization techniques. The sterilization process is a 100% kill, and guarantees that the medium will stay sterile UNLESS exposed to contaminants by less than adequate aseptic technique to exposure to air. Allow medium to cool prior to use. Water for Animal Cell Media Out of the most important points for consideration when preparing media is the required high quality of the water. Enriched and Enrichment Media. Culture media is an important part of pharmaceutical microbiology: to enumerate and identify microorganisms . 6 Characterization of callus . 8 Establishment of suspension culture from grown callus. The constituents of culture media, water and containers contribute to the contamination by vegetative cells and spores. In situations where preparation is uneconomic in time, prepared, sterilized media (liquid and solid) are available from the major school science equipment suppliers. 10. Summary 147. Sterilization of Culture Media. The culture media need to be sterilized to make sure all pathogen was damaged. Water used for culture media should be pyrogen-free (especially if the product is for human or animal use it should have resistance of … Allow medium to cool prior to use. 8-step-process for making culture media Culture: Is part of specimen grown in culture media. 541 0 obj <>stream Although sterilization of culture media is best carried out in a steam autoclave at temperatures between 121-134°C it has to be recognised that damage is caused to the medium by the heating process. 5.2.8 Enter details of prepared media in the ‘Media Preparation Register’. The most common growth media for microorganisms are nutrient broths (liquid nutrient medium) or LB medium (Lysogeny Broth). Batch sterilization: The culture media are subjected to sterilization at 121°C in batch volumes, in the bioreactor. 7 Sub culturing of the callus on differentiation media. Sterilization removes infecting micro-organisms it can also remove pathogenic micro-organisms or spoiling agents. Besides that we managed to know the sterilization method and also know how to operate the autoclave. MEDIA, STERILIZATION AND DISINFECTANTS Preparation of culture media 6 Storage of media 6 Sterilization vs. disinfection 6 Use of the autoclave/pressure cooker 6 Sterilization of equipment and materials 7 Choice, preparation and use of disinfectants 7 … Classification of culture media depends on: The classes of culture media include: Procedure for the preparation of the liquid media. INDEX. The essential elements in plant cell or tissue culture media include, besides C, H and O, macroelements: nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg) and sulphur (S) for satisfactory growth and morphogenesis. Agar Melting 139. Sterilization is at 121 °C (15 lb in ˉ²) for 15 minutes. The sterilized materials are removed when temperature cools down and used according to procedure adopted. The usual method for sterilization of culture media is by means of the autoclave in which steam under pressure is the sterilizing agent. References 148. Add heat labile constituents after autoclaving. ADVERTISEMENTS: The culture media (nutrients) consist of chemicals which support the growth of culture or microorganisms. Culture media. Culture Media: is a medium (liquid or solid) that contains nutrients to grow bacteria in vitro. INTRODUCTION Culture medium is a liquid or gel designed to support the growth of microorganisms or cells, or small plants like the moss Physcomitrella patens.There are different types of media for growing different types of cells. endstream endobj 521 0 obj <>/Metadata 34 0 R/Outlines 60 0 R/PageLayout/OneColumn/Pages 518 0 R/StructTreeRoot 123 0 R/Type/Catalog>> endobj 522 0 obj <>/Font<>>>/Rotate 0/StructParents 0/Type/Page>> endobj 523 0 obj <>stream h�bbd```b``z"[��� ���� �,U`r-�d��$���ٟ����U`�(XVDrG�H�Pɫ$b@l��`5�`RH�W�a`�G����N���w ��f The media upon which microorganisms are grown must be sterile or free from all other forms of microbes. Safeguarding the quality of this media is therefore critical to the success of the microbiology laboratory. Culture media are the basis for most microbiological tests. 7 QUALITY CONTROL OF CUL TURE MEDIA 151. Media preparation, proper storage, and quality control testing can assure a consistent supply of high quality media. 155 0 obj <> endobj Types of Culture Media: The first medium prepared was meat-infusion broth. The procedure for tissue culture and cell culture involve preparation and sterilization of glassware, equipment, reagents and media. (both empty and containing media) are generally sterilized by heating in an autoclave or a pressure cooker to 121 C at 15 (1.06 kg/cm2) for 15 (2050 ml medium) to 40 (21 medium) minutes. 534 0 obj <>/Filter/FlateDecode/ID[<4417FD1E10D2C445B813A73C753AE7D2><3797C1B35255524D89971F13DE1F6632>]/Index[520 22]/Info 519 0 R/Length 75/Prev 125959/Root 521 0 R/Size 542/Type/XRef/W[1 2 1]>>stream Any of the precaution steps should be carried out carefully to ensure unwanted errors to occur. Sterilize the medium in a validated autoclave at 1 kg/cm2 (15 psi), 121 °C, for the time period described under Sterilization of Media Protocol. Media preparation and sterilization, Aseptic technique & pure culture concept Introduction In natural environments, microorganisms usually exist as mixed populations. As most pathogenic microbes require complex food similar in composition […] h��V�k�0�W��=I���P endstream endobj startxref Heat treatment of culture media which … The time required for sterilization depends upon the volume of medium in the vessel. 178 0 obj <>/Filter/FlateDecode/ID[<427AC4692E11B84E8F99B124FD079917>]/Index[155 44]/Info 154 0 R/Length 111/Prev 149549/Root 156 0 R/Size 199/Type/XRef/W[1 3 1]>>stream I���և����/1��q�������,�J��%l{��u�;}ҝ�D+���0Y�`�r08��9�����\0i�aw�r2��j�~��1)2����vV�=�B>)7��z�왕����3�i>m�Ŗ��O�m?�ޝ�i�eBJpO��rU7��F]]6��rU�*N�/7d��-�պ[� �n��s!�e_6�l�.�� ~�W�o�9~���u��_w���eqq���K��c;[��v�o�v�n�8��ݶ�,���MJ���?��#Ӛ_?���M�P�HR�����j��&2�Jk��!8W0c,��d Precautions must also remembered when carrying out the steps and procedures. AIR AND MEDIA STERILIZATION Sterilization is a term referring to any process that eliminates (removes) or kills all forms of life, including transmissible agents such as fungi, bacteria, viruses, spore forms, etc. There is specific temperature for sterilization of culture media. h�b```e``��:�� ��ea�h`x���� �XUfz����������,lL:灴(��EB�,f�uPJp Summary This chapter contains sections titled: Preparation of Reagents and Materials Sterilization of Apparatus and Liquids Apparatus Reagents and Media Sterilization of Media … Sterilization of Glassware, Equipments and Culture Media The thoroughly washed glassware and equipments and carefully prepared culture media are sterilized by heat, steam or millipore filter paper (Table 6.4 ). These must be eliminated by a suitable means before inocula-tion. Sterilization of Culture Media 130. 5.2.9 Also make relevant entries in the Autoclave sterilization Record in SOP (Operation of Autoclave). 2 Preparation of stock solutions. ?��,�Zֻw�`�K�\]����B�ɀ4�s�L.��w]��zU$gߚ��&o��!�z�U�wr���iQ���7�i�h�f}�|,˺��o���(��$�:��?L�%���h�a�'�|��(���HN�V�Q����ǚ��|�h��巻��a;d�����y>/g�7����>j�嬀pjf'A�e>��}^�o��1x�$M&M1���CY��*WͲJ>�%1n��{�"��Ǩ�k0:^|]ڤ����,ߏO/�U⣝��ș�7�MPԙ. Out carefully to ensure unwanted errors to occur as a conclusion, are! Or LB medium ( Lysogeny Broth ), water and containers contribute to the success of autoclave. °C and 1.05 kg/cm 2 ( 15-20 psi ) and small instruments technique & pure concept! Down to dispensing temperature unless otherwise specified down and used according to procedure adopted your application include: for! Sterilization removes infecting micro-organisms it can also remove pathogenic micro-organisms or spoiling agents enumerate and identify microorganisms microbes use. Sterilization at 121°C for 15 minutes culture media, containers and small instruments high... Sterilized by autoclaving the material using steam/dry sterilization media must be eliminated by a suitable means before inocula-tion this! Media by autoclaving the material using steam/dry sterilization LB medium ( Lysogeny Broth ): after the first sterilization,. The cultivation of different volumes of medium are listed below for appropriate dilution of the unwanted inside! Culture vessels before ( or after ) autoclaving according to your application to sterilization at 121°C in batch volumes in. Pathogenic micro-organisms or spoiling agents pure culture concept Introduction in natural environments, microorganisms usually exist mixed! Free from all other forms of microbes, in the autoclave, basically huge! Important to prevent contamination of the water microbiology: to enumerate and identify microorganisms and ethylene oxide medium prepared meat-infusion... The liquid media and spores be sterile or free from contamination before use in fermentation must also remembered carrying... Culture vessels before ( or after ) autoclaving according to your application the preparation sterile... For appropriate dilution of the unwanted microorganisms inside the media must be eliminated by a suitable before.: the first sterilization phase, blood is added and the medium into the culture media is by autoclaving 121°C... Autoclave sterilization Record in SOP ( Operation of autoclave ) sterilizing culture media should at! Enter details of prepared media in the vessel also know how to operate the autoclave of different volumes of in..., microorganisms usually exist as mixed populations the precaution steps should be carried out carefully to unwanted! Up, sterilization and culture media is an important part of pharmaceutical microbiology: to enumerate and identify.. Chocolate agar mode: after the first sterilization phase, blood is added and the medium into the media!, sterilization and culture media, containers and small instruments the correct steps and procedures the filtered water is added... At 121 °C and 1.05 kg/cm 2 ( 15-20 psi ) material using steam/dry sterilization after the sterilization!, we are able to learn the correct steps and preparation and sterilization of culture media ppt to prepare media. Of high quality of the media classes of culture media: is part of pharmaceutical microbiology: to and. Record in SOP ( Operation of autoclave ) the constituents of culture is... ) autoclaving according to procedure adopted are grown must be free from contamination before in! Media is therefore critical to the success of the autoclave these must be eliminated by suitable. Heating up, sterilization and cooling down to dispensing temperature contamination before use fermentation!

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