Sep 09, 2020

Public workspaceTPGY BROTH (TRYPTONE PEPTONE GLUCOSE YEAST EXTRACT BROTH)

This protocol is a draft, published without a DOI.
  • 1Soil and Water Research Infrastructure
  • Anaerobic and Molecular Microbiology Lab, Biology Centre CAS
    Tech. support email: eva.petrova@bc.cas.cz
Ana Lara
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External link: http://www.himedialabs.com/TD/M969.pdf
Protocol Citation: Roey Angel, Ana Lara-Rodriguez, Eva Petrova 2020. TPGY BROTH (TRYPTONE PEPTONE GLUCOSE YEAST EXTRACT BROTH). protocols.io https://protocols.io/view/tpgy-broth-tryptone-peptone-glucose-yeast-extract-3e5gjg6
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 28, 2019
Last Modified: September 09, 2020
Protocol Integer ID: 23741
Abstract
For the enrichment of Clostridium sp. from soil samples, sufficient for 100 cultivation tubes.
Guidelines
Materials
MATERIALS
ReagentPeptone A FROM MEATBio Basic Inc.Catalog #G213.SIZE.500g
ReagentSodium thioglycolateBio Basic Inc.Catalog #S0265.SIZE.25g
ReagentBD Bacto™ Yeast Extract Becton Dickinson (BD)Catalog #212750
ReagentEnzymatic Digest Casein HydrolysateCatalog #AAJ12855Q1
ReagentDextroseMerck MilliporeSigma (Sigma-Aldrich)Catalog #D9434
Safety warnings
Use heat reasistant gloves to take media out of the autoclave and to put and take out the media from the water bath.
Before start
1. Make sure that the glass tubes are sterile.
2. Pour media only in the flow box.
Medium preparation
Medium preparation

  1. Dilute in 1000 ml distilled water:

Amount50 g Casein Hydrolysate
Amount5 g Peptic Digest from Animal Tissue (Peptone A)
Amount20 g Yeast Extract
Amount4 g Dextrose
Amount1 g Sodium Thioglycolate

Final pH ( at 25°C) 7.0±0.2

Note
Heat if necessary, to dissolve the medium completely.

Sterilize by autoclaving at 15 lbs pressure Temperature121 °C for Duration00:15:00 .
Refrigerate Temperature4 °C the sterile medium until use.
Before inoculation of media
Before inoculation of media

Pour Amount10 mL of media in sterile tubes.
Heat media in water bath Temperature100 °C Duration00:10:00 OR alternative do a gass exchange using the gassing station for 3 minutes to each tube OR put tubes in to the anaerobic tent for Duration24:00:00 to Duration48:00:00 before inoculating.