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Difco Nutrient Agar is a rich medium commercially available from BD and routinely used for cultivating many non-fastidious organisms. It contains beef extract and peptones; exact composition will vary between batches. | Difco Nutrient Agar is a rich medium commercially available from BD and routinely used for cultivating many non-fastidious organisms. It contains beef extract and peptones; exact composition will vary between batches. | ||
Many Streptomyces sp. grow well on DNA, but developmental and/or antibiotic production phenotypes may differ compared to growth on other media (e.g. [[SFM]]). | Many ''Streptomyces'' sp. grow well on DNA, but developmental and/or antibiotic production phenotypes may differ compared to growth on other media (e.g. [[SFM]]). | ||
==Preparation== | ==Preparation== | ||
| Line 9: | Line 9: | ||
*4g Difco Nutrient Broth Powder | *4g Difco Nutrient Broth Powder | ||
*10g Agar | *10g Agar | ||
*1L | *1L dH<sub>2</sub>O | ||
'''Instructions''' | '''Instructions''' | ||
#For solid medium, add 0.8g of DNB powder into each 250ml conical flask | #For solid medium, add 0.8g of DNB powder into each 250ml conical flask | ||
#Add 2g of Agar | #Add 2g of Agar | ||
#Add 200ml | #Add 200ml dH<sub>2</sub>O | ||
#Autoclave | #Autoclave | ||
==Uses== | ==Uses== | ||
*[[ | *''Streptomyces'' Colony PCR (see [[High GC PCR]] for instructions) | ||
*[[Spore titres]] | *[[Spore titres]] | ||
*[[Re-isolation experiments]] | *[[Re-isolation experiments]] | ||
*[[Selection for resistance to salt-sensitive antibiotics]] | *[[Selection for resistance to salt-sensitive antibiotics]] | ||
*[[Streptomyces Bacteriophage plaque assay]] | |||
==Notes== | ==Notes== | ||
For growth in liquid culture, [[DNB]] can be made without the agar. | For growth in liquid culture, [[DNB]] can be made without the agar. | ||
According to the recipe in Practical Streptomyces Genetics, it should be 8g of DNB per litre, however some groups choose to use 4g instead. This has little effect on the ability of ''Streptomyces'' to grow on the soild medium, [[DNA]]. | |||
==References== | ==References== | ||
Latest revision as of 12:50, 5 May 2020
Difco Nutrient Agar (DNA)
Difco Nutrient Agar is a rich medium commercially available from BD and routinely used for cultivating many non-fastidious organisms. It contains beef extract and peptones; exact composition will vary between batches.
Many Streptomyces sp. grow well on DNA, but developmental and/or antibiotic production phenotypes may differ compared to growth on other media (e.g. SFM).
Preparation
Per Litre:
- 4g Difco Nutrient Broth Powder
- 10g Agar
- 1L dH2O
Instructions
- For solid medium, add 0.8g of DNB powder into each 250ml conical flask
- Add 2g of Agar
- Add 200ml dH2O
- Autoclave
Uses
- Streptomyces Colony PCR (see High GC PCR for instructions)
- Spore titres
- Re-isolation experiments
- Selection for resistance to salt-sensitive antibiotics
- Streptomyces Bacteriophage plaque assay
Notes
For growth in liquid culture, DNB can be made without the agar.
According to the recipe in Practical Streptomyces Genetics, it should be 8g of DNB per litre, however some groups choose to use 4g instead. This has little effect on the ability of Streptomyces to grow on the soild medium, DNA.
