Actinobacteria isolation from marine environments - Revision history

Laia: /* Media selection */

2019-08-30T15:06:01Z

Media selection

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 Adjust to pH to 7.0 - 7.5 and supplement with nalidixic acid (10 μg ml−1) and nystatin (50 μg ml−1) to inhibit the growth of Gram-negative bacteria and fungi.
 ==References==
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 - Bredholt, H. et al. (2008) ‘Actinomycetes from sediments in the Trondheim fjord, Norway: Diversity and biological activity’, Marine Drugs. doi: 10.3390/md6010012.

Laia: /* Pre-treatment methods of marine samples */

2019-08-30T14:59:18Z

Pre-treatment methods of marine samples

← Older revision		Revision as of 14:59, 30 August 2019
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	==Applying high frequency radiation==		==Applying high frequency radiation==
	Apply superhigh frequency radiation directly to sediment samples by placing the sediment solution in a Eppendorf tube and microwave it for 45 seconds at 2460 MHz and 80 W (4).		Apply superhigh frequency radiation directly to sediment samples by placing the sediment solution in a Eppendorf tube and microwave it for 45 seconds at 2460 MHz and 80 W (4).

			==Dilute / Heat==
			Add dry sediment to 3 mL of seawater (dilutions 1:3 or 1:6), heat it to 55°C for 6 min, and inoculate 50-70 µL of the resulting suspension onto an agar plate (2).

	=Media selection=		=Media selection=

Laia: /* Media selection */

2019-08-30T13:50:58Z

Media selection

← Older revision		Revision as of 13:50, 30 August 2019
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	=Media selection=		=Media selection=
	Soil agar:		Soil agar (4):

	Filtered soil extract (mix 200 g of soil in 1 L of water and boil for 30 min) – 1 L		Filtered soil extract (mix 200 g of soil in 1 L of water and boil for 30 min) – 1 L

	Agar – 20 g		Agar – 20 g
	~~Adjust to pH – 7.0~~


Line 28:		Line 29:


	Modified organic agar 2 Gause:		Modified organic agar 2 Gause (4):

	Peptone – 5.0 g		Peptone – 5.0 g

	Trypton – 3.0 g		Trypton – 3.0 g

	Glucose – 10 g		Glucose – 10 g

	NaCl – 5.0 g		NaCl – 5.0 g

	Tap water – 1 L		Tap water – 1 L


	Modified organic agar 2 Gause with 3% sea salt added		Modified organic agar 2 Gause with 3% sea salt added (4)


	Mineral agar 1 Gause:		Mineral agar 1 Gause (4):

	Starch‐soluble – 20.0 g		Starch‐soluble – 20.0 g

	K2HPO4– 0.5 g		K2HPO4– 0.5 g

	MgSO4– 0.5 g		MgSO4– 0.5 g

	KNO3– 1.0 g		KNO3– 1.0 g

	NaCl – 0.5 g		NaCl – 0.5 g

	FeSO4−0.01 g		FeSO4−0.01 g

	Agar – 20.0 g		Agar – 20.0 g

	Tap water – 1 L		Tap water – 1 L

	Soy‐bean meal agar:
			Soy‐bean meal agar (4):

	Soy‐bean meal – 3.0 g		Soy‐bean meal – 3.0 g

	KNO3– 0.2 g		KNO3– 0.2 g

	K2HPO4– 0.5 g		K2HPO4– 0.5 g

	MgSO4– 0.4 g		MgSO4– 0.4 g

	Agar 20.0 g		Agar 20.0 g

	Tap water – 1 L		Tap water – 1 L

	~~Soy‐bean meal agar with 3% sea salt added~~

			Soy‐bean meal agar with 3% sea salt added (4)


			Yeast‐corn‐starch agar (4):

	~~Yeast‐corn‐starch agar~~
	Starch‐soluble – 10.0 g		Starch‐soluble – 10.0 g

	Yeast extract – 10 g		Yeast extract – 10 g

	Corn meal extract – 10.0 g		Corn meal extract – 10.0 g
	NaCl – 2.0 g~~, agar~~ – 20.0 g
			NaCl – 2.0 g

			Agar – 20.0 g

	Tap water – 1 L		Tap water – 1 L


	Pea‐meal agar:		Pea‐meal agar (4):

	Pea meal – 10.0 g		Pea meal – 10.0 g

	Glucose – 10.0 g		Glucose – 10.0 g

	NaCl – 5.0 g		NaCl – 5.0 g

	CaCO3– 1.0 g		CaCO3– 1.0 g

	Agar – 20 g		Agar – 20 g

	Tap water – 1.0 L		Tap water – 1.0 L

Laia: /* Media selection */

2019-08-30T13:46:16Z

Media selection

← Older revision		Revision as of 13:46, 30 August 2019
Line 19:		Line 19:

	=Media selection=		=Media selection=
			Soil agar:
			Filtered soil extract (mix 200 g of soil in 1 L of water and boil for 30 min) – 1 L
			Agar – 20 g
			Adjust to pH – 7.0


			Soil agar with 3% sea salt added


			Modified organic agar 2 Gause:
			Peptone – 5.0 g
			Trypton – 3.0 g
			Glucose – 10 g
			NaCl – 5.0 g
			Tap water – 1 L


			Modified organic agar 2 Gause with 3% sea salt added


			Mineral agar 1 Gause:
			Starch‐soluble – 20.0 g
			K2HPO4– 0.5 g
			MgSO4– 0.5 g
			KNO3– 1.0 g
			NaCl – 0.5 g
			FeSO4−0.01 g
			Agar – 20.0 g
			Tap water – 1 L

			Soy‐bean meal agar:
			Soy‐bean meal – 3.0 g
			KNO3– 0.2 g
			K2HPO4– 0.5 g
			MgSO4– 0.4 g
			Agar 20.0 g
			Tap water – 1 L

			Soy‐bean meal agar with 3% sea salt added


			Yeast‐corn‐starch agar
			Starch‐soluble – 10.0 g
			Yeast extract – 10 g
			Corn meal extract – 10.0 g
			NaCl – 2.0 g, agar – 20.0 g
			Tap water – 1 L


			Pea‐meal agar:
			Pea meal – 10.0 g
			Glucose – 10.0 g
			NaCl – 5.0 g
			CaCO3– 1.0 g
			Agar – 20 g
			Tap water – 1.0 L

			Adjust to pH to 7.0 - 7.5 and supplement with nalidixic acid (10 μg ml−1) and nystatin (50 μg ml−1) to inhibit the growth of Gram-negative bacteria and fungi.


	==References==		==References==

Laia at 16:04, 26 July 2019

2019-07-26T16:04:09Z

← Older revision		Revision as of 16:04, 26 July 2019
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	# Drying sediment overnight in a laminar flow hood until clumping occurs. Press a sterile foam plug against the sediment sample and onto the surface of an agar plate making a serial dilution (2).		# Drying sediment overnight in a laminar flow hood until clumping occurs. Press a sterile foam plug against the sediment sample and onto the surface of an agar plate making a serial dilution (2).

	#(2) Drying sediment overnight in a laminar flow hood until clumping occurs. Dilute dried sediment with 5 ml of sterile seawater and mix well. Add 50 µl of the solution onto an agar plate and spread with an spreader (2).		(2#) Drying sediment overnight in a laminar flow hood until clumping occurs. Dilute dried sediment with 5 ml of sterile seawater and mix well. Add 50 µl of the solution onto an agar plate and spread with an spreader (2).

	==Mixing the samples with seawater==		==Mixing the samples with seawater==

Laia at 16:03, 26 July 2019

2019-07-26T16:03:39Z

← Older revision		Revision as of 16:03, 26 July 2019
Line 7:		Line 7:
	# Drying sediment overnight in a laminar flow hood until clumping occurs. Press a sterile foam plug against the sediment sample and onto the surface of an agar plate making a serial dilution (2).		# Drying sediment overnight in a laminar flow hood until clumping occurs. Press a sterile foam plug against the sediment sample and onto the surface of an agar plate making a serial dilution (2).

	## Drying sediment overnight in a laminar flow hood until clumping occurs. Dilute dried sediment with 5 ml of sterile seawater and mix well. Add 50 µl of the solution onto an agar plate and spread with an spreader (2).		#(2) Drying sediment overnight in a laminar flow hood until clumping occurs. Dilute dried sediment with 5 ml of sterile seawater and mix well. Add 50 µl of the solution onto an agar plate and spread with an spreader (2).

	==Mixing the samples with seawater==		==Mixing the samples with seawater==

Laia: /* Drying marine samples with laminar air flow hood */

2019-07-26T16:03:10Z

Drying marine samples with laminar air flow hood

← Older revision		Revision as of 16:03, 26 July 2019
Line 7:		Line 7:
	# Drying sediment overnight in a laminar flow hood until clumping occurs. Press a sterile foam plug against the sediment sample and onto the surface of an agar plate making a serial dilution (2).		# Drying sediment overnight in a laminar flow hood until clumping occurs. Press a sterile foam plug against the sediment sample and onto the surface of an agar plate making a serial dilution (2).

	# Drying sediment overnight in a laminar flow hood until clumping occurs. Dilute dried sediment with 5 ml of sterile seawater and mix well. Add 50 µl of the solution onto an agar plate and spread with an spreader (2).		## Drying sediment overnight in a laminar flow hood until clumping occurs. Dilute dried sediment with 5 ml of sterile seawater and mix well. Add 50 µl of the solution onto an agar plate and spread with an spreader (2).

	==Mixing the samples with seawater==		==Mixing the samples with seawater==

Laia at 16:02, 26 July 2019

2019-07-26T16:02:04Z

← Older revision		Revision as of 16:02, 26 July 2019
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	Several drying pre-treatment methods can be used to enhance the isolation of Actinobacteria from marine environments (2):		Several drying pre-treatment methods can be used to enhance the isolation of Actinobacteria from marine environments (2):

	1- Drying sediment overnight in a laminar flow hood until clumping occurs. Press a sterile foam plug against the sediment sample and onto the surface of an agar plate making a serial dilution (2).		# Drying sediment overnight in a laminar flow hood until clumping occurs. Press a sterile foam plug against the sediment sample and onto the surface of an agar plate making a serial dilution (2).

	2- Drying sediment overnight in a laminar flow hood until clumping occurs. Dilute dried sediment with 5 ml of sterile seawater and mix well. Add 50 µl of the solution onto an agar plate and spread with an spreader (2).		# Drying sediment overnight in a laminar flow hood until clumping occurs. Dilute dried sediment with 5 ml of sterile seawater and mix well. Add 50 µl of the solution onto an agar plate and spread with an spreader (2).

	==Mixing the samples with seawater==		==Mixing the samples with seawater==

Laia at 15:59, 26 July 2019

2019-07-26T15:59:40Z

← Older revision		Revision as of 15:59, 26 July 2019
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	Apply superhigh frequency radiation directly to sediment samples by placing the sediment solution in a Eppendorf tube and microwave it for 45 seconds at 2460 MHz and 80 W (4).		Apply superhigh frequency radiation directly to sediment samples by placing the sediment solution in a Eppendorf tube and microwave it for 45 seconds at 2460 MHz and 80 W (4).

	==Media selection==		=Media selection=

	==References==		==References==

Laia: Laia moved page Category:Actinobacteria isolation from marine environments to Actinobacteria isolation from marine environments over redirect

2019-07-26T15:57:12Z

Laia moved page Category:Actinobacteria isolation from marine environments to Actinobacteria isolation from marine environments over redirect

← Older revision	Revision as of 15:57, 26 July 2019
(No difference)