Mesoplama florum:Culture media - Revision history

Tk at 04:34, 14 November 2010

Tk — Sun, 14 Nov 2010 04:34:00 GMT

←Older revision		Revision as of 04:34, 14 November 2010
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	\| 50 ml		\| 50 ml
	\|}		\|}
-	[[Category:Material]] ~~and~~ [[Category:Standard Media]][[Category:Mesoplasma florum]]	+	[[Category:Material]][[Category:Standard Media]][[Category:Mesoplasma florum]]

Torsten Waldminghaus at 08:54, 31 October 2008

Torsten Waldminghaus — Fri, 31 Oct 2008 08:54:05 GMT

←Older revision		Revision as of 08:54, 31 October 2008
Line 96:		Line 96:
	\| 50 ml		\| 50 ml
	\|}		\|}
-		+	[[Category:Material]] and [[Category:Standard Media]][[Category:Mesoplasma florum]]
-	[[Category:Material]] [[Category:Mesoplasma florum]]	+

Tk: /* ATCC 1161 Medium */

Tk — Sat, 10 Nov 2007 22:05:57 GMT

ATCC 1161 Medium

←Older revision		Revision as of 22:05, 10 November 2007
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	Thaw Sigma H-1138 heat inactivated horse serum. For agar, bring both horse serum and centrifuged yeast extract to 37C. Horse serum at 55C will sediment protein.		Thaw Sigma H-1138 heat inactivated horse serum. For agar, bring both horse serum and centrifuged yeast extract to 37C. Horse serum at 55C will sediment protein.

-	Assemble ~~150ml~~ bottle top 0.22μ filter on the 1 liter bottle. For agar, it is helpful to put the bottle on a foam insulating pad to delay cooling. Add by vacuum filtration:	+	Assemble 500 ml bottle top 0.22μ filter on the 1 liter bottle. For agar, it is helpful to put the bottle on a foam insulating pad to delay cooling. Add by vacuum filtration:

	{\|		{\|

Reshma P. Shetty at 20:11, 8 May 2007

Reshma P. Shetty — Tue, 08 May 2007 20:11:38 GMT

←Older revision		Revision as of 20:11, 8 May 2007
Line 96:		Line 96:
	\| 50 ml		\| 50 ml
	\|}		\|}
		+
		+	[[Category:Material]] [[Category:Mesoplasma florum]]

Tk at 21:10, 14 November 2006

Tk — Tue, 14 Nov 2006 21:10:01 GMT

←Older revision		Revision as of 21:10, 14 November 2006
Line 38:		Line 38:
	\| 400 μl		\| 400 μl
	\|-		\|-
-	\| 2% phenol red	+	\| 2% phenol red or 1% cresol red
-	\| 2 ml	+	\| 2 ml (5 ml for cresol red)
	\|}		\|}

-	Adjust pH to 7.4 with ~~100 mM~~ HCl. Measured pH of as-made broth is 7.46, probably close enough.	+	Adjust pH to 7.4 with 1 M HCl or 1 M NaOH. Measured pH of as-made broth is 7.46, probably close enough. For cresol red containing solution, adjust pH to 7.8

-	Phenol red 2% solution must be neutralized with sufficient NaOH to convert it to the sodium salt. The phenol red amount can be adjusted to control the depth of color to taste.	+	Phenol red 2% solution must be neutralized with sufficient NaOH to convert it to the sodium salt. The phenol or cresol red amount can be adjusted to control the depth of color to taste.

	====SP-4 (ATCC 988) Medium====		====SP-4 (ATCC 988) Medium====

Kathmc at 13:28, 9 December 2005

Kathmc — Fri, 09 Dec 2005 13:28:34 GMT

New page

====ATCC 1161 Medium====

Mix a base broth in a 1 liter 45mm cap bottle the following chemicals:

{|
|-
| Heart Infusion Broth (Difco 0038)
| 17.5 g
|-
| Sucrose
| 40 g
|-
| Agar Noble (Difco 0142) if necessary
| 12 g
|-
| DI Water
| 710 ml
|-
|}

Autoclave at 121C for 20 minutes and cool to room temperture (broth) or 55C (Agar) on a 55C water bath.

Centrifuge 2 x 45 ml of Gibco 18180-059 liquid yeast extract at high speed to remove sediment. Pour only 45 ml into centrifuge bottles, eliminating as much as possible of the sediment in the bottom of the Gibco stock.

Thaw Sigma H-1138 heat inactivated horse serum. For agar, bring both horse serum and centrifuged yeast extract to 37C. Horse serum at 55C will sediment protein.

Assemble 150ml bottle top 0.22μ filter on the 1 liter bottle. For agar, it is helpful to put the bottle on a foam insulating pad to delay cooling. Add by vacuum filtration:

{|
|-
| Horse serum (Sigma H1138)
| 200 ml
|-
| Yeast extract
| 90 ml
|-
| Penicillin G 500,000 U/ml
| 400 μl
|-
| 2% phenol red
| 2 ml
|}

Adjust pH to 7.4 with 100 mM HCl. Measured pH of as-made broth is 7.46, probably close enough.

Phenol red 2% solution must be neutralized with sufficient NaOH to convert it to the sodium salt. The phenol red amount can be adjusted to control the depth of color to taste.

====SP-4 (ATCC 988) Medium====

Mix a base broth containing:

{|
|-
| Mycoplasma broth base (BBL 111458)
| 3.5 g
|-
| Tryptone
| 10 g
|-
| Peptone
| 5.3 g
|-
| Glucose
| 5.0 g
|-
| DI water
| 615 ml
|}

Adjust pH to 7.5 and autoclave at 121C for 20 minutes.

Centrifuge 2 x 17.5 ml of liquid yeast extract (Gibco 18180-059) at high speed to remove sediment.

Mix 2 g of Yeastolate (Difco 5577) in 100 ml DI water.

Sterile filter into the cooled medium:

{|
|-
| penicillin G 500,000 U/ml
| 400 μl
|-
| 2% phenol red
| 2 ml
|-
| 2% Yeastolate (Difco 5577)
| 100 ml
|-
| Newborn calf serum
| 170 ml
|-
| liquid yeast extract
| 35 ml
|-
| 10x CMRL-1066 solution
| 50 ml
|}