[Skip to Content](https://www.sigmaaldrich.com#main-content) [![MilliporeSigma](https://www.sigmaaldrich.com/static/logos/purple/millipore_sigma.svg)](https://www.sigmaaldrich.com/US/en) Products Cart0 USEN Products ProductsApplicationsServicesResourcesSupport [Login / Register](https://www.sigmaaldrich.com/oidc-sign-in) [Order Lookup](https://www.sigmaaldrich.com/US/en/order-lookup) [Quick Order](https://www.sigmaaldrich.com/US/en/quick-order) Cart0 [Home](https://www.sigmaaldrich.com/US/en)[Media Preparation](https://www.sigmaaldrich.com/US/en/applications/cell-culture-and-cell-culture-analysis/cell-culture-by-technique/cell-culture-media-preparation)Media Sterilization – Plant Tissue Culture Protocol # Media Sterilization – Plant Tissue Culture Protocol Plant tissue culture media are generally sterilized by autoclaving at 121 °C and 1.05 kg/cm2 (15-20 psi). The time required for sterilization depends upon the volume of medium in the vessel. The minimum times required for sterilization of different volumes of medium are listed below. It is advisable to dispense medium in small aliquots whenever possible as many media components are broken down on prolonged exposure to heat. There is evidence that medium exposed to temperatures in excess of 121 °C may not properly gel or may result in poor cell growth. ## Minimum Autoclaving Time for Plant Tissue Culture Medium | | | |----------------------------------|----------------------------| | Volume of Medium per Vessel (mL) | Minimum Autoclaving (min)* | | 25 | 20 | | 50 | 25 | | 100 | 28 | | 250 | 31 | | 500 | 35 | | 1000 | 40 | | 2000 | 48 | | 4000 | 63 | \*Minimum autoclaving time includes the time required for the liquid volume to reach the sterilizing temerature (121 °C) and 15 min. at 121 °C (Burger, 1988). Times may vary due to differences in autoclaves. Validation with your system is recommended. Several medium components are considered thermolabile and should not be autoclaved. Stock solutions of the heat labile components are prepared and filter sterilized through a 0.22 µm filter into a sterile container. The filtered solution is aseptically added to the culture medium, which has been autoclaved and allowed to cool to approximately 35-45 °C. The medium is then dispensed under sterile conditions. Experimentation with your system is recommended. __Related Articles__ - [Cell Culture Media Preparation Using MilliShot™ Single Dose Antibiotics](https://www.sigmaaldrich.com/US/en/technical-documents/protocol/cell-culture-and-cell-culture-analysis/cell-culture-media-preparation/cell-culture-media-preparation-millishot-single-dose-antibiotics) - [Protocol for Charcoal-stripping FBS to Deplete Hormones](https://www.sigmaaldrich.com/US/en/technical-documents/protocol/cell-culture-and-cell-culture-analysis/cell-culture-media-preparation/charcoal-stripped-heat-inactivated-fbs-protocols) - [Freezing and Thawing FBS and Other Sera](https://www.sigmaaldrich.com/US/en/technical-documents/protocol/cell-culture-and-cell-culture-analysis/cell-culture-media-preparation/storing-and-thawing-serum) Top __Sign In To Continue__ To continue reading please sign in or create an account. Sign In__Don't Have An Account?__Register Support[Customer Support](https://www.sigmaaldrich.com/US/en/support/customer-support)[Contact Us](https://www.sigmaaldrich.com/US/en/collections/offices)[FAQ](https://maestro.my.site.com/knowledgeportal/s/)[Safety Data Sheets (SDS)](https://www.sigmaaldrich.com/US/en/documents-search?tab=sds)[Certificates (COA/COO)](https://www.sigmaaldrich.com/US/en/documents-search?tab=coa)[Quality & Regulatory](https://www.sigmaaldrich.com/US/en/life-science/quality-and-regulatory-management)[Calculators & Apps](https://www.sigmaaldrich.com/US/en/support/calculators-and-apps)[Webinars](https://www.sigmaaldrich.com/US/en/collections/webinars) Orders[Order Lookup](https://www.sigmaaldrich.com/US/en/order-lookup)[Quick Order](https://www.sigmaaldrich.com/US/en/quick-order)[Custom Products](https://www.sigmaaldrich.com/US/en/services/custom-products)[eCommerce Solutions](https://www.sigmaaldrich.com/US/en/life-science/ecommerce/ecommerce-solutions) Company[About Us](https://www.sigmaaldrich.com/US/en/life-science/about-us)[Responsibility](https://www.sigmaaldrich.com/US/en/life-science/ssbi)[Events](https://www.sigmaaldrich.com/US/en/collections/events)[Press Releases](https://www.sigmaaldrich.com/US/en/collections/press)[Programs](https://www.sigmaaldrich.com/US/en/life-science/partnership-programs)[Careers](https://careers.merckgroup.com/global/en)[Offices](https://www.sigmaaldrich.com/US/en/collections/offices) Social Media [![LinkedIn icon](https://www.sigmaaldrich.com/content/dam/cms-commons/sigmaaldrich/marketing/global/images/icons/linkedin.svg)](https://www.linkedin.com/company/merck-life-science) [![X icon](https://www.sigmaaldrich.com/content/dam/cms-commons/sigmaaldrich/marketing/global/images/icons/x-icon.svg)](https://x.com/Merck_lifesci) [![Facebook Icon](https://www.sigmaaldrich.com/content/dam/cms-commons/sigmaaldrich/marketing/global/images/icons/facebook.svg)](https://www.facebook.com/Merck.lifescience) [![Instagram Icon](https://www.sigmaaldrich.com/content/dam/cms-commons/sigmaaldrich/marketing/global/images/icons/instagram.svg)](https://www.instagram.com/mercklifescience/) [![YouTube Icon](https://www.sigmaaldrich.com/content/dam/cms-commons/sigmaaldrich/marketing/global/images/icons/youtube.svg)](https://www.youtube.com/@MerckLifeScience/featured) MilliporeSigma __Research. Development. Production.__ We are a leading supplier to the global Life Science industry with solutions and services for research, biotechnology development and production, and pharmaceutical drug therapy development and production. [![Vibrant M string](https://www.sigmaaldrich.com/content/dam/cms-commons/sigmaaldrich/marketing/global/images/icons/vibrant-m-string-1-rrvcrp.svg)](https://www.sigmaaldrich.com/US/en) [Sigma-Aldrich® Solutions](https://www.sigmaaldrich.com/US/en/life-science/sigma-aldrich)[BioReliance® Solutions](https://www.sigmaaldrich.com/US/en/life-science/bioreliance)[Millipore® Solutions](https://www.sigmaaldrich.com/US/en/life-science/millipore)[SAFC® Solutions](https://www.sigmaaldrich.com/US/en/life-science/safc)[Milli-Q® Solutions](https://www.sigmaaldrich.com/US/en/life-science/milliq)[Supelco® Solutions](https://www.sigmaaldrich.com/US/en/life-science/supelco) © 2026 Merck KGaA, Darmstadt, Germany and/or its affiliates. All Rights Reserved, including Text and Data Mining for AI training and similar technologies. Reproduction of any materials from the site is strictly forbidden without permission. [Site Use Terms](https://www.sigmaaldrich.com/US/en/life-science/legal/site-use-terms)|[Privacy Policy](https://www.sigmaaldrich.com/US/en/life-science/legal/privacy-statement)|[General Terms and Conditions of Sale](https://www.sigmaaldrich.com/US/en/life-science/legal/terms-and-conditions)|[Copyright Consent](https://www.sigmaaldrich.com/US/en/life-science/legal/copyright-consent) |[Site Map](https://www.sigmaaldrich.com/US/en/site-map) An unknown error has occured.