Part 4: Fertilization and Development - Revision history

imported>Virgilio: moved Part 4:Fertilization and Development to Part 4: Fertilization and Development

2011-11-16T15:20:00Z

moved Part 4:Fertilization and Development to Part 4: Fertilization and Development

← Older revision	Revision as of 15:20, 16 November 2011
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136.159.234.84 at 22:42, 14 January 2010

2010-01-14T22:42:50Z

← Older revision		Revision as of 22:42, 14 January 2010
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			Protocol submitted by Janet Heasman [http://www.xenbase.org/community/person.do?method=display&personId=733&tabId=0]


	'''Fertilize embryos'''		'''Fertilize embryos'''

imported>Xenbase: /* Related Articles */

2009-12-22T00:08:26Z

← Older revision		Revision as of 00:08, 22 December 2009
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	==Related Articles==		==Related Articles==
			*[[Oocyte Transfer Technique]]
	*[[Part 1: Choosing Oligodeoxyribonucleotides (ODNs)]]		*[[Part 1: Choosing Oligodeoxyribonucleotides (ODNs)]]
	*[[Part 2: ODN Modification]]		*[[Part 2: ODN Modification]]
	*[[Part 3: Host Transfer Technique]]		*[[Part 3: Host Transfer Technique]]

imported>Xenbase at 00:06, 22 December 2009

2009-12-22T00:06:59Z

← Older revision		Revision as of 00:06, 22 December 2009
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	As it is easier to distinguish the 5 colors from each other early in development rather than later, the embryos are subdivided into separate dishes at this time and are monitored carefully through development to observe differences between the control and experimental embryos. In addition, embryos may be fixed and frozen for further analysis at later stages.		As it is easier to distinguish the 5 colors from each other early in development rather than later, the embryos are subdivided into separate dishes at this time and are monitored carefully through development to observe differences between the control and experimental embryos. In addition, embryos may be fixed and frozen for further analysis at later stages.

			==Related Articles==
			*[[Part 1: Choosing Oligodeoxyribonucleotides (ODNs)]]
			*[[Part 2: ODN Modification]]
			*[[Part 3: Host Transfer Technique]]

imported>Xenbase at 00:05, 22 December 2009

2009-12-22T00:05:01Z

← Older revision		Revision as of 00:05, 22 December 2009
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	'''Fertilize embryos'''		'''Fertilize embryos'''

	Three hours after the oocytes are transferred to the host frog, the frog is squeezed to lay eggs at 20-30 minute intervals and fertilized with a sperm suspension, as per a normal in vitro fertilization (Holwill et al., 1987). The percentage of experimental oocytes that are laid by the female varies from one experiment to the next, but as long as the female does not stop laying eggs most of the dyed oocytes can be recovered. Of these only a portion of them will fertilize, and a 20-40% rate of fertilization is usual for transferred and uninjected oocytes. Usually a female will give 3-4 good fertilizations per experiment.		Three hours after the oocytes are transferred to the host frog, the frog is squeezed to lay eggs at 20-30 minute intervals and fertilized with a sperm suspension, as per a normal in vitro fertilization (Holwill et al., 1987). The percentage of experimental oocytes that are laid by the female varies from one experiment to the next, but as long as the female does not stop laying eggs most of the dyed oocytes can be recovered. Of these only a portion of them will fertilize, and a 20-40% rate of fertilization is usual for transferred and uninjected oocytes. Usually a female will give 3-4 good fertilizations per experiment.

	'''Dejelly embryos'''		'''Dejelly embryos'''

	Embryos are dejellied in 2% cysteine pH 7.8 in 0.1X MMR (Appendix A) after the first cell cycle. Briefly, they are gently swirled until the embryos lie close together instead of being held apart by the jelly coat. Then embryos are washed three times in a large excess volume of 0.1X MMR. Both host and transferred embryos are then allowed to develop in 0.1XMMR.		Embryos are dejellied in 2% cysteine pH 7.8 in 0.1X MMR (Appendix A) after the first cell cycle. Briefly, they are gently swirled until the embryos lie close together instead of being held apart by the jelly coat. Then embryos are washed three times in a large excess volume of 0.1X MMR. Both host and transferred embryos are then allowed to develop in 0.1XMMR.

	As it is easier to distinguish the 5 colors from each other early in development rather than later, the embryos are subdivided into separate dishes at this time and are monitored carefully through development to observe differences between the control and experimental embryos. In addition, embryos may be fixed and frozen for further analysis at later stages.		As it is easier to distinguish the 5 colors from each other early in development rather than later, the embryos are subdivided into separate dishes at this time and are monitored carefully through development to observe differences between the control and experimental embryos. In addition, embryos may be fixed and frozen for further analysis at later stages.

imported>Xenbase: Created page with ''''Fertilize embryos''' Three hours after the oocytes are transferred to the host frog, the frog is squeezed to lay eggs at 20-30 minute intervals and fertilized with a sperm sus…'

2009-12-22T00:04:48Z

Created page with ''''Fertilize embryos''' Three hours after the oocytes are transferred to the host frog, the frog is squeezed to lay eggs at 20-30 minute intervals and fertilized with a sperm sus…'

New page

'''Fertilize embryos'''
Three hours after the oocytes are transferred to the host frog, the frog is squeezed to lay eggs at 20-30 minute intervals and fertilized with a sperm suspension, as per a normal in vitro fertilization (Holwill et al., 1987). The percentage of experimental oocytes that are laid by the female varies from one experiment to the next, but as long as the female does not stop laying eggs most of the dyed oocytes can be recovered. Of these only a portion of them will fertilize, and a 20-40% rate of fertilization is usual for transferred and uninjected oocytes. Usually a female will give 3-4 good fertilizations per experiment.

'''Dejelly embryos'''
Embryos are dejellied in 2% cysteine pH 7.8 in 0.1X MMR (Appendix A) after the first cell cycle. Briefly, they are gently swirled until the embryos lie close together instead of being held apart by the jelly coat. Then embryos are washed three times in a large excess volume of 0.1X MMR. Both host and transferred embryos are then allowed to develop in 0.1XMMR.

As it is easier to distinguish the 5 colors from each other early in development rather than later, the embryos are subdivided into separate dishes at this time and are monitored carefully through development to observe differences between the control and experimental embryos. In addition, embryos may be fixed and frozen for further analysis at later stages.