May 02, 2022

Public workspaceCBM for propagation of in vitro plantlets

This protocol is a draft, published without a DOI.
  • 1Realizing Increased Photosynthetic Efficiency (RIPE)
Lynn Doran
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Protocol Citation: Lynn Doran, Amanda P. De Souza 2022. CBM for propagation of in vitro plantlets. protocols.io https://protocols.io/view/cbm-for-propagation-of-in-vitro-plantlets-b8kfrutn
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it’s working
Created: May 02, 2022
Last Modified: July 11, 2023
Protocol Integer ID: 61799
Keywords: Cassava, tissue culture, media, propagation
Funders Acknowledgement:
Realizing Increased Photosynthetic Efficiency (RIPE) that is funded by the Bill & Melinda Gates Foundation, Foundation for Food and Agriculture Research, and the U.K. Foreign, Commonwealth & Development Office
Grant ID: OPP1172157
Abstract
Media preparation for in vitro propagation of plantlets. Validated for cassava in tissue culture.
Attachments
Icon for MS media for cassava propagation.docx
MS media for cassava...
15KB
Guidelines
  • Thoroughly sterilize all materials and work surfaces before pouring media.
  • Use sterile technique after autoclaving, when pouring media into containers.
  • Wear a lab coat and clean gloves that have been thoroughly soaked in 70% ethanol during media pouring.
  • Do not turn off laminar flow hood blower fans during media setting time.
  • Do not leave UV light on once media is in the flow hood. The UV light will damage the vitamin content of the media.
Materials
Reagents
  • Copper Sulfate
  • Murashige & Skoog Basal Medium with Vitamins (Phytotech Labs M519)
  • Sucrose
  • Carbencillin, C500
  • Sodium Hydroxide, 1 M
  • 70% Ethanol for sterilization

Materials
  • CONTAINER FOR PLANT TISSUE CULTURE, WITH LID, 330 ML, PS, STERILE (Greiner Bio-one 968161)
  • Weigh dishes or paper
  • Autoclave tape

Equipment
  • Spatulas
  • Graduated cylinder, 1000 mL
  • Beakers, 2000 mL
  • Reagent bottle, 2000 mL
  • Pipette, 20-200 ul
  • Pipette, 1-5 mL
  • Spray bottle for 70% Ethanol
  • Analytical Balance
  • pH Meter
  • Autoclave bin
  • Autoclave
  • Laminar Flow Hood
Safety warnings
Autoclaves generate steam under high pressure. Read all safety guidelines and receive proper training before operating.