ActinoBase - User contributions [en-gb]

Protocols

2019-09-26T15:42:40Z

Tommclean:

[[File:Actinobase-icons-trans_Protocols.png|150px|Protocols|link=Protocols]]

[[Characterisation of Actinomycetes]]

[[Cloning, Transformation and Expression of Gene Clusters]]

[[CRISPR/Cas9-mediated genome editing]]

[[General Growth and Maintenance of Streptomyces sp.]]

[[GUS assay]] (β-Glucuronidase assay)

[[High GC PCR]]

[[Isolating Actinomycetes from the environment]]

[[Metabolite extractions]]

[[Microscopy]]

[[Mutagenesis]]

[[Nucleic Acid Methods]]

[[Protein Methods]]

Media recipes

2019-09-26T15:38:37Z

Tommclean:

{{DISPLAYTITLE:Media Recipes}}

[[File:Actinobase-icons-trans_Media-recipes.png|150px|Media Recipes|link=Media_recipes]]

=Media Recipes=

[[A1]]

[[CM]] (Complete medium)

[[DNA]] (Difco Nutrient Agar) | [[DNB]] (Difco Nutrient Broth) | [[SNA]] (Soft Nutrient Agar)

[[Emerson medium]]

[[Gause's No. 1]]

[[GYM]] (Glucose + Yeast Extract + Malt Extract)

[[HT]] (Hickey-Tresner)

[[HV]]

[[IMA]] (Smash)

[[LB]] (Lysogeny Broth) | [[Lennox]] (Lennox agar) | [[Luria]] (Luria agar)

[[MC agar]] (Milk Casein Agar)

[[MM]] (Minimal medium)

[[SFM | MS]] (Mannitol Soya agar - a.k.a. Soya Flour Mannitol (SFM))

[[MYM]] (Maltose + Yeast Extract + Malt Extract)

[[NMMP]] (minimal liquid medium)

[[OT]] (oligotrophic agar)

[[PDA]] (Potato Dextrose Agar)

[[R2]] | [[R2YE]] | [[R5]]

[[SC]] (Starch Casein agar)

[[SFM]] (Soya Flour Mannitol - a.k.a. Mannitol Soya agar)

[[SMM]] (supplemented liquid minimal medium) | [[SMMS]] (supplemented minimal medium, solid)

[[SPY]] (Salt Peptone Yeast)

[[TSB]] (Tryptone Soy Broth)

[[YEME]] (Yeast Extract + Malt Extract)

[[YMPD]] (Yeast Meat Peptone Dextrose)

[[YP]] (Yeast Peptone) | [[YPD]] (Yeast Peptone Dextrose)

[[2X YT]]

====International ''Streptomyces'' Project Media (ISP's)====

[[ISP2]]

[[ISP3]]

[[ISP4]]

[[ISP5]]

[[ISP6]]

[[ISP7]]

==Media Supplements==

[[Antibiotic stocks and working concentrations]]

[[Indicator compounds]]

[[Media supplements for activating cryptic biosynthetic gene clusters]]

[[Media supplements for induction gene expression from inducible promoters]]

[[Trace element solution]]

Lennox

2019-09-26T15:26:28Z

Tommclean:

=Lennox Broth=
Known as Lennox Broth (or sometimes, mistakenly, as Luria-Bertani medium), this rich medium is used routinely for growth of Escherichia coli. There are several variants on this medium, with differing amounts of NaCl; glucose may also be added, as in the original recipe (Bertani 1951).

==Preparation==
Per litre:
*5g Yeast Extract
*5g NaCl
*10g Tryptone
*Up to 1L dH2O

'''Instructions'''
#Add 5g Yeast Extract, 5g NaCl and 10g Tryptone to a beaker
#Add 800ml dH2O and stir until all solids are fully suspended
#Make up to 1L with dH2O
#For solid media, add 4g agar per 200ml LB
#Autoclave

===-NaCl Lennox===
Per litre:
*5g Yeast Extract
*10g Tryptone
*Up to 1L dH2O

==Uses==
*Routine growth of Escherichia coli
*Checking for contamination in Streptomyces spore stocks or cultures

==Notes==
This is the Lennox formulation, with 5g/L NaCl. Alternatively, 10g/L NaCl can be used (Miller formulation) or no salt (-NaCl).
The -NaCl (no salt) version of Lennox should be used when using salt sensitive antibiotics like [[hygromycin]]: for example, when cloning into [[pSS170]]. (Alternatively, [[DNA]] or [[DNB]] can be used for hygromycin selection.)

To make liquid Lennox medium, omit the agar and autoclave as above.

==References==
Lennox, E. S. 1955. Transduction of linked genetic characters of the host by bacteriophage P1.
Virology. 1:190-206.

Luria

2019-09-26T15:25:58Z

Tommclean: Created page with "=Luria Broth= Known as Luria Broth (or sometimes, mistakenly, as LB or Lennox medium), this rich medium is used routinely for growth of Escherichia coli. There are se..."

=Luria Broth=
Known as Luria Broth (or sometimes, mistakenly, as LB or Lennox medium), this rich medium is used routinely for growth of Escherichia coli. There are several variants on this medium, with differing amounts of NaCl; glucose may also be added, as in the original recipe (Bertani 1951).

==Preparation==
Per litre:
*5g Yeast Extract
*0.5g NaCl
*10g Tryptone
*Up to 1L dH2O

'''Instructions'''
#Add 5g Yeast Extract, 0.5g NaCl and 10g Tryptone to a beaker
#Add 800ml dH2O and stir until all solids are fully suspended
#Make up to 1L with dH2O
#For solid media, add 4g agar per 200ml LB
#Autoclave

===-NaCl Lennox===
Per litre:
*5g Yeast Extract
*10g Tryptone
*Up to 1L dH2O

==Uses==
*Routine growth of Escherichia coli
*Checking for contamination in Streptomyces spore stocks or cultures

==Notes==
This is the Luria formulation, with 0.5g/L NaCl. Alternatively, 10g/L NaCl can be used (Miller formulation) or no salt (-NaCl).
The -NaCl (no salt) version of Luria should be used when using salt sensitive antibiotics like [[hygromycin]]: for example, when cloning into [[pSS170]]. (Alternatively, [[DNA]] or [[DNB]] can be used for hygromycin selection.)

To make liquid Luria medium, omit the agar and autoclave as above.

==References==
Luria, S. E., and J. W. Burrous. 1955. Hybridization between Escherichia coli and Shigella. J.
Bacteriol. 74:461-476.

Lennox

2019-09-26T15:23:45Z

Tommclean: Created page with "=Lennox Broth= Known as Lennox Broth (or sometimes, mistakenly, as Luria-Bertani medium), this rich medium is used routinely for growth of Escherichia coli. There are..."

=Lennox Broth=
Known as Lennox Broth (or sometimes, mistakenly, as Luria-Bertani medium), this rich medium is used routinely for growth of Escherichia coli. There are several variants on this medium, with differing amounts of NaCl; glucose may also be added, as in the original recipe (Bertani 1951).

==Preparation==
Per litre:
*5g Yeast Extract
*5g NaCl
*10g Tryptone
*Up to 1L dH2O

'''Instructions'''
#Add 5g Yeast Extract, 5g NaCl and 10g Tryptone to a beaker
#Add 800ml dH2O and stir until all solids are fully suspended
#Make up to 1L with dH2O
#For solid media, add 4g agar per 200ml LB
#Autoclave

===-NaCl Lennox===
Per litre:
*5g Yeast Extract
*10g Tryptone
*Up to 1L dH2O

==Uses==
*Routine growth of Escherichia coli
*Checking for contamination in Streptomyces spore stocks or cultures

==Notes==
This is the Lennox formulation, with 5g/L NaCl. Alternatively, 10g/L NaCl can be used (Miller formulation) or no salt (-NaCl).
The -NaCl (no salt) version of Lennox should be used when using salt sensitive antibiotics like [[hygromycin]]: for example, when cloning into [[pSS170]]. (Alternatively, [[DNA]] or [[DNB]] can be used for hygromycin selection.)

To make liquid LB medium, omit the agar and autoclave as above.

==References==
Lennox, E. S. 1955. Transduction of linked genetic characters of the host by bacteriophage P1.
Virology. 1:190-206.

Media recipes

2019-09-26T15:19:39Z

Tommclean:

R5

2019-09-26T15:12:56Z

Tommclean:

=R5=

==Preparation==
Make up the following solution:
*103 g Sucrose
*0.25 g K2SO4
*10.12 g MgCl2.6H2O
*10 g Glucose
*0.1 g Difco Casaminoacids
*2 ml [[Trace element solution]]
*5 g Difco yeast extract
*5.73 g TES buffer
*To 1000 ml dH2O

Pour 100 ml of the solution into 250 ml Erlenmeyer flasks each containing 2.2 g Difco Bacto agar.
Sterilise by autoclaving.
At the time of use, remelt the medium and add to each flask the following autoclaved solutions in the order listed:
*1 ml KH2PO4 (0.5%)
*0.4 ml CaCl2.2H2O (5M, 3.68%)
*1.5 ml L-proline (20%)
*0.7 ml NaOH (1N) (unsterilised is okay)
*0.75 ml Required growth factors for auxotrophs

==Uses==
*See [[R2YE]] - just an alternate way of making but folklore suggests less reproducible results on R5.

==Notes==
''[[Streptomyces clavuligerus]]'' does not utilise glucose and it must be substituted by glycerol.

==References==

Kieser, T., Bibb, M.J., Buttner, M.J., Chater, K.F. and Hopwood, D.A. (2000) Practical Streptomyces Genetics: John Innes Foundation, Norwich Research Park, Colney, Norwich NR4 7UH, UK

R5

2019-09-26T15:12:39Z

Tommclean:

=R5=

==Preparation==
Make up the following solution:
*103 g Sucrose
*0.25 g K2SO4
*10.12 g MgCl2.6H2O
*10 g Glucose
*0.1 g Difco Casaminoacids
*2 ml [[trace element solution]]
*5 g Difco yeast extract
*5.73 g TES buffer
*To 1000 ml dH2O

Pour 100 ml of the solution into 250 ml Erlenmeyer flasks each containing 2.2 g Difco Bacto agar.
Sterilise by autoclaving.
At the time of use, remelt the medium and add to each flask the following autoclaved solutions in the order listed:
*1 ml KH2PO4 (0.5%)
*0.4 ml CaCl2.2H2O (5M, 3.68%)
*1.5 ml L-proline (20%)
*0.7 ml NaOH (1N) (unsterilised is okay)
*0.75 ml Required growth factors for auxotrophs

==Uses==
*See [[R2YE]] - just an alternate way of making but folklore suggests less reproducible results on R5.

==Notes==
''[[Streptomyces clavuligerus]]'' does not utilise glucose and it must be substituted by glycerol.

==References==

Kieser, T., Bibb, M.J., Buttner, M.J., Chater, K.F. and Hopwood, D.A. (2000) Practical Streptomyces Genetics: John Innes Foundation, Norwich Research Park, Colney, Norwich NR4 7UH, UK

R5

2019-09-26T15:12:07Z

Tommclean: Created page with "=R5= ==Preparation== Make up the following solution: *103 g Sucrose *0.25 g K2SO4 *10.12 g MgCl2.6H2O *10 g Glucose *0.1 g Difco..."

=R5=

==Preparation==
Make up the following solution:
*103 g Sucrose
*0.25 g K2SO4
*10.12 g MgCl2.6H2O
*10 g Glucose
*0.1 g Difco Casaminoacids
*2 ml [[trace elements solution]]
*5 g Difco yeast extract
*5.73 g TES buffer
*To 1000 ml dH2O

Pour 100 ml of the solution into 250 ml Erlenmeyer flasks each containing 2.2 g Difco Bacto agar.
Sterilise by autoclaving.
At the time of use, remelt the medium and add to each flask the following autoclaved solutions in the order listed:
*1 ml KH2PO4 (0.5%)
*0.4 ml CaCl2.2H2O (5M, 3.68%)
*1.5 ml L-proline (20%)
*0.7 ml NaOH (1N) (unsterilised is okay)
*0.75 ml Required growth factors for auxotrophs

==Uses==
*See [[R2YE]] - just an alternate way of making but folklore suggests less reproducible results on R5.

==Notes==
''[[Streptomyces clavuligerus]]'' does not utilise glucose and it must be substituted by glycerol.

==References==

Kieser, T., Bibb, M.J., Buttner, M.J., Chater, K.F. and Hopwood, D.A. (2000) Practical Streptomyces Genetics: John Innes Foundation, Norwich Research Park, Colney, Norwich NR4 7UH, UK

MYM

2019-09-26T15:07:23Z

Tommclean:

=Maltose Yeast Extract Medium (MYM)=

Maltose-Yeast Extract-Malt Extract (MYM) medium is a rich medium used especially for the growth of ''[[Streptomyces venezuelae]]'', as it sporulates to >90% completion in liquid MYM [2]. Other Streptomyces sp. may also grow well on this medium.

==Preparation==
Per litre:
*4g Maltose
*4g Yeast Extract Oxoid
*10g Malt Extract
*500ml Tap Water
*500ml dH2O

'''Instructions'''

#Dissolve all ingredients in 500ml tap water & 250ml dH2O
#pH to 7.2-7.3 with NaOH
#add dH2O up to 1L
#For solid medium, add 2g agar & 100mL liquid MYM to each flask
#Autoclave, expect a precipitate to form afterward.
#For optimal ''[[Streptomyces venezuelae]]'' sporulation, add [[Trace element solution]] at a 1:500 dilution.

==Uses==
*Routine growth of ''[[Streptomyces venezuelae]]''
*Developmental studies on ''[[Streptomyces venezuelae]]''

==Notes==
If using for growth measurements in liquid medium, it is best to allow the precipitate to settle overnight before pipetting the medium into growth vessels - take from the top to avoid the precipitate.

==References==
1. Stuttard C. 1982. Temperate phages of Streptomyces venezuelae: lysogeny and host specificity shown by phages SV1 and SV2. Microbiology 128:115–121.

2. Bibb MJ, Domonkos A, Chandra G, Buttner MJ. 2012. Expression of the chaplin and rodlin hydrophobic sheath proteins in Streptomyces venezuelae is controlled by σBldN and a cognate anti-sigma factor, RsbN. Mol. Microbiol. 84:1033–1049

MediaWiki:Sidebar

2019-09-12T15:36:43Z

Tommclean:

MediaWiki:Sidebar

2019-09-12T15:33:09Z

Tommclean:

* navigation
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* Menu
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MediaWiki:Sidebar

2019-09-12T15:32:04Z

Tommclean:

* navigation
** helppage|help-mediawiki
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** randompage-url|randompage

* Menu
** background|Background
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** Plasmids|Plasmids
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** genomes|Genomes
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** groups|Research Groups
** Admin_contact_page|Contact
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** Links|Links
** suppliers|Suppliers

* SEARCH
* TOOLBOX
* LANGUAGES

*Twitter
<h3>Join the conversation on twitter!</h3>
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Exploration

2019-08-29T15:57:42Z

Tommclean:

==General overview==

Exploration is an alternative developmental stage of ''Actinobacteria'' consisting of rapid horizontal hyphal growth on [[YP]] agar, or [[YPD]] agar in the presence of certain fungi, with little to no branching. The exploring colony will also produce volatile organic compounds which induce exploration in distal colonies. Not every Actinobacteria is capable of exploring but was first described in ''Streptomyces venezuelae'' along with others [1].

Useful host strains:
*Streptomyces venezuelae [1]
This species is currently the most well characterised for exploration.

Abiotic induction protocol:
*Make 33-50mL [[YP]] plates
*Inoculate 5μL of bacterial spore stock in the centre of the plate
*Incubate at 30ºC for 14 days
*Image plate

This protocol is more reliable as you are not reliant on the activity of the yeast co-culture

Biotic induction protocol:
*Grow ''Saccharomyces cerevisiae'' in an overnight culture of liquid [[YPD]]
*Make 33-50mL [[YPD]] plates
*Inoculate 5μL of bacterial spore stock in the centre of the plates
*Inoculate 5μL of ''Saccharomyces cerevisiae'' overnight culture next to the other inoculation and blend gently
*Incubate at 30ºC for 14 days
*Image plate

This protocol can be less reliable due to the nature of the yeast co-culture but allows for reciprocal testing if required.

References:
[1] Jones, S., Ho, L., Rees, C., Hill, J., Nodwell, J. and Elliot, M. (2017). Streptomyces exploration is triggered by fungal interactions and volatile signals. eLife, 6.

Protocol adapted for Actinobase by Thomas C. McLean, The University of East Anglia

Exploration

2019-08-29T15:35:16Z

Tommclean: Created page with "==General overview== [1] Useful host strains: *Streptomyces venezuelae [1] This species is currently the most well character..."

==General overview==

[1]

Useful host strains:
*Streptomyces venezuelae [1]
This species is currently the most well characterised for exploration.

Abiotic induction protocol:
*Make 33-50mL [[YP]] plates
*Inoculate 5μL of ''Actinomyces spp..'' spore stock in the centre of the plate
*Incubate at 30ºC for 14 days
*Image plate

This protocol is more reliable as you are not reliant on the activity of the yeast co-culture

Biotic induction protocol:
*Grow ''Saccharomyces cerevisiae'' in an overnight culture of liquid [[YPD]]
*Make 33-50mL [[YPD]] plates
*Inoculate 5μL of ''Actinomyces spp.'' spore stock in the centre of the plates
*Inoculate 5μL of ''Saccharomyces cerevisiae'' overnight culture next to the other inoculation and blend gently
*Incubate at 30ºC for 14 days
*Image plate

This protocol can be less reliable due to the nature of the yeast co-culture but allows for reciprocal testing if required.

References:
[1] Jones, S., Ho, L., Rees, C., Hill, J., Nodwell, J. and Elliot, M. (2017). Streptomyces exploration is triggered by fungal interactions and volatile signals. eLife, 6.

Protocol adapted for Actinobase by Thomas C. McLean, The University of East Anglia

Characterisation of Actinomycetes

2019-08-29T15:13:35Z

Tommclean: Created page with "Exploration Production of microbial volatile organic compounds"

[[Exploration]]

[[Production of microbial volatile organic compounds]]

Protocols

2019-08-29T15:12:26Z

Tommclean:

[[File:Actinobase-icons-trans_Protocols.png|150px|Protocols|link=Protocols]]

[[Characterisation of Actinomycetes]]

[[Cloning, Transformation and Expression of Gene Clusters]]

[[CRISPR/Cas9-mediated genome editing]]

[[General Growth and Maintenance of Streptomyces sp.]]

[[High GC PCR]]

[[Mutagenesis]]

[[Nucleic Acid Methods]]

[[Protein Methods]]

[[Microscopy]]

[[Isolating Actinomycetes from the environment]]

Protocols

2019-08-29T15:08:10Z

Tommclean:

[[File:Actinobase-icons-trans_Protocols.png|150px|Protocols|link=Protocols]]

[[Cloning, Transformation and Expression of Gene Clusters]]

[[CRISPR/Cas9-mediated genome editing]]

[[Exploration]]

[[General Growth and Maintenance of Streptomyces sp.]]

[[High GC PCR]]

[[Mutagenesis]]

[[Nucleic Acid Methods]]

[[Protein Methods]]

[[Microscopy]]

[[Isolating Actinomycetes from the environment]]

Recruitment

2019-08-02T12:45:46Z

Tommclean: Created page with "=Recruitment= Please feel free to post information and links to job opportunities etc below: *Please also remember to remove the links once filled."

=Recruitment=
Please feel free to post information and links to job opportunities etc below:
*Please also remember to remove the links once filled.

MediaWiki:Sidebar

2019-08-02T12:43:52Z

Tommclean:

SMMS

2019-08-01T15:57:57Z

Tommclean: Created page with "=SMMS (Supplemented liquid minimal media, solid)= SMM is a solid supplemented minimal medium suitable for growth of many ''Streptomyces'' sp. ==Preparation== Per litre: *81.9..."

=SMMS (Supplemented liquid minimal media, solid)=
SMM is a solid supplemented minimal medium suitable for growth of many ''Streptomyces'' sp.

==Preparation==
Per litre:
*81.9 mL PEG 6000 (BDH 6.1% w/v in distilled water)
*2.5 mL Mg2SO4.7H2O - 5mM final
*10 mL TES bugger (0.25M, pH 7.2) - 25mM final
*1 mL NaH2PO4 + K2HPO4 (50mM each) - 1mM final
*2 mL glucose (50% w/v)
*1 mL Antifoam 289 (Sigma A 5551)(1% w/v)
*0.1 mL Trace element solution
*1 mL Difco Casaminoacids (20% w/v)
*100 mL Total
*25 ml/L Glycine (0.5% final)

Trace element solution:
0.1 g/L of the following:
*ZnSO4.7H2O
*FeSO4.7H2O
*MnCl2.4H2O
*CaCl2.6H2O
*NaCl
Make fresh every 2-4 weeks and filter-sterilise. Store at 4º

'''Instructions'''
#With the exception of the trace element solution each solution must be made up and autoclaved separately
#The solutions should be mixed in the given order and then made up to 1L with sterile distilled water
#Addition of 10 g/L agar

==References==
Strauch E, Takano E, Baylts H, Bibb M. The stringent response in ''Streptomyces coelicolor'' A3(2). ''Molecular Microbiology''. 1991;5(2):289-298.

SMM

2019-08-01T15:52:02Z

Tommclean: /* SMM (Supplemented liquid minimal media) */

=SMM (Supplemented liquid minimal media)=
SMM is a supplemented minimal medium suitable for growth of many ''Streptomyces'' sp.

==Preparation==
Per litre:
*81.9 mL PEG 6000 (BDH 6.1% w/v in distilled water)
*2.5 mL Mg2SO4.7H2O - 5mM final
*10 mL TES bugger (0.25M, pH 7.2) - 25mM final
*1 mL NaH2PO4 + K2HPO4 (50mM each) - 1mM final
*2 mL glucose (50% w/v)
*1 mL Antifoam 289 (Sigma A 5551)(1% w/v)
*0.1 mL Trace element solution
*1 mL Difco Casaminoacids (20% w/v)
*100 mL Total
*25 ml/L Glycine (0.5% final)

Trace element solution:
0.1 g/L of the following:
*ZnSO4.7H2O
*FeSO4.7H2O
*MnCl2.4H2O
*CaCl2.6H2O
*NaCl
Make fresh every 2-4 weeks and filter-sterilise. Store at 4º

'''Instructions'''
#With the exception of the trace element solution each solution must be made up and autoclaved separately
#The solutions should be mixed in the given order and then made up to 1L with sterile distilled water

==References==
Strauch E, Takano E, Baylts H, Bibb M. The stringent response in ''Streptomyces coelicolor'' A3(2). ''Molecular Microbiology''. 1991;5(2):289-298.

SMM

2019-08-01T15:42:07Z

Tommclean:

=SMM (Supplemented liquid minimal media)=
Strauch et al., 19911

==Preparation==
Per litre:
*81.9 mL PEG 6000 (BDH 6.1% w/v in distilled water)
*2.5 mL Mg2SO4.7H2O - 5mM final
*10 mL TES bugger (0.25M, pH 7.2) - 25mM final
*1 mL NaH2PO4 + K2HPO4 (50mM each) - 1mM final
*2 mL glucose (50% w/v)
*1 mL Antifoam 289 (Sigma A 5551)(1% w/v)
*0.1 mL Trace element solution
*1 mL Difco Casaminoacids (20% w/v)
*100 mL Total
*25 ml/L Glycine (0.5% final)

Trace element solution:
0.1 g/L of the following:
*ZnSO4.7H2O
*FeSO4.7H2O
*MnCl2.4H2O
*CaCl2.6H2O
*NaCl
Make fresh every 2-4 weeks and filter-sterilise. Store at 4º

'''Instructions'''
#With the exception of the trace element solution each solution must be made up and autoclaved separately
#The solutions should be mixed in the given order and then made up to 1L with sterile distilled water

SMM

2019-08-01T15:30:50Z

Tommclean: Created page with "=SMM (Supplemented liquid minimal media)= Strauch et al., 19911 ==Preparation== Per litre: *81.9 mL PEG 6000 (BDH 6.1% w/v in distilled water) *2.5 mL Mg2</su..."

=SMM (Supplemented liquid minimal media)=
Strauch et al., 19911

==Preparation==
Per litre:
*81.9 mL PEG 6000 (BDH 6.1% w/v in distilled water)
*2.5 mL Mg2SO4.7H2O - 5mM final
*10 mL TES bugger (0.25M, pH 7.2) - 25mM final
*1mL NaH2PO4 + K2HPO4 (50mM each) - 1mM final
*2 mL glucose (50% w/v)
*1 mL Antifoam 289 (Sigma A 5551)(1% w/v)
*0.1 mL Trace element solution
*1 mL Difco Casaminoacids (20% w/v)
*100 mL Total
*25 ml/L Glycine (0.5% final)

'''Instructions'''
#Dissolve all of the ingredients except the agar and the glucose in the distilled water
#Adjust the pH to 7.0-7.2
#Dispense 200ml into flasks each containing 2g agar
#Autoclave
#After autoclaving, add 4 ml sterile 50% glucose solution (autoclaved separately or filter-sterilized) to each flask
#Add any other growth factors required for auxotrophic mutants

YP

2019-07-04T15:50:54Z

Tommclean:

For YP please see [[YPD]]. Ignore part about the addition of dextrose.

YP

2019-07-04T15:50:08Z

Tommclean: Created page with "See YPD"

See [[YPD]]

YPD

2019-07-04T15:42:12Z

Tommclean: Created page with "=Yeast Peptone Dextrose= Yeast Peptone Dextrose (YPD) medium is composed of Yeast Peptone medium (YP) with the addition of dextrose (D-glucose). YPD is a general medium used..."

=Yeast Peptone Dextrose=

Yeast Peptone Dextrose (YPD) medium is composed of Yeast Peptone medium (YP) with the addition of dextrose (D-glucose). YPD is a general medium used for the cultivation of fungal species and investigating ''Streptomyces'' exploration [1].

==Preparation==
Per Litre:
*10g yeast extract
*20g bacto-peptone
*20g Agar
*1L with dH20 water

20% dextrose:
*20g dextrose
*100ml dH20
*Filter sterilise

'''Instructions'''
#Dissolve 10g yeast extract and 20g bacto-peptone in 1L dH20 making YP solution
#Add 2g agar to each 250mL flask
#Add 100mL YP solution to each flask
#Autoclave - this makes YP agar
#When cool add 10mL 20% dextrose solution to 100mL media

==Uses==
*Explorer phenotype investigation
*Fungal cultivation

==Notes==
*YP has been shown to induce exploration in ''Streptomyces venezuelae'' whilst YPD has been shown to repress it unless co-cultivated with certain fungal strains

==References==
1. Jones S, Ho L, Rees C, Hill J, Nodwell J, Elliot M. ''Streptomyces'' exploration is triggered by fungal interactions and volatile signals. ''eLife''. 2017;6.