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Cytostatic Factor (CSF) Extract Preparation for Spindle Assembly Protocol - http://www.bio.com/protocolstools/protocol.jhtml?id=p1422

Category: Cell Biology and Cell Culture: Cell Cycle Protocols: Mitosis Protocols

Cytostatic factor extract (CSF) preparation for spindle assembly protocol. Protocol includes tips such as: The quality of the eggs is essential for good CSF extracts. Always sacrifice quantity for quality when trying to make functional extracts. - [Read Cytostatic Factor (CSF) Extract Preparation for Spindle Assembly Protocol]

Cytostatic Factor (CSF) Extract Preparation for Xenopus Spindle Assembly Protocol - http://www.bio.com/protocolstools/protocol.jhtml?id=p1422

Category: Xenopus Protocols: Xenopus Egg Extracts Protocols

Protocol for cytostatic factor extract preparation for spindle assembly. The quality of the eggs is essential for good CSF extracts. Always sacrifice quantity for quality when trying to make functional extracts. Discard any batches of eggs that have "puff balls" or activated eggs that constitute more than 10% of the eggs. Use laid eggs and collect eggs at about 16 to 17 hours after priming with Progesterone (found in Pregnant Mare Serum Gonadotropin (PMSG)). - [Read Cytostatic Factor (CSF) Extract Preparation for Xenopus Spindle Assembly Protocol]

De-Jellying Eggs Protocol - http://tropicalis.berkeley.edu/home/manipulate_embryos/dejelly.html

Category: Xenopus Protocols: Xenopus Embryo Manipulation Protocols

Protocol for de-jellying eggs. - [Read De-Jellying Eggs Protocol]

Differences in Injection of X. laevis and X. tropicalis - http://tropicalis.berkeley.edu/home/manipulate_embryos/microinjection/differences.html

Category: Xenopus Protocols: Xenopus Injection Protocols

Differences in injection of X. laevis and X. tropicalis. Includes: X. tropicalis lays eggs about 4 hours after a boost of hCG; In vitro fertilization is not nearly as efficient in trops compared to laevis; X. tropicalis embryos are much softer than X. laevis embryos; X. tropicalis embryos whose jelly coats are removed by cysteine have a loose sticky vitelline membrane; X. tropicalis do not have a "summer slump". - [Read Differences in Injection of X. laevis and X. tropicalis]

Fixation and Permeabilization of C. elegans Protocol - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=35

Category: C. elegans Protocols: C. elegans Fixing Protocols

Protocol for fixation and permeabilization of C. elegans. Protocol works for all stages except dauers (which won't open) and hypoclorite-treated eggs (which disintegrate). Luckily, hypochlorite treatment and fixation are by themselves sufficient to open eggs. - [Read Fixation and Permeabilization of C. elegans Protocol]

Genetically-Engineered Mouse Models - http://www.bcm.edu/dtmc/

Category: Microinjection Protocols: Microinjection of DNA Protocols

Microinjection Services – mouse ES cells into blastocysts and DNA into pronucleus of fertilized eggs. - [Read Genetically-Engineered Mouse Models]

Liquid Culture of Worms Protocol - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=22

Category: C. elegans Protocols: C. elegans Cell Culture Protocols

Protocol for liquid culture of worms. Includes: superbroth; S- basal; worm plates; GROWING THE BACTERIA (WORM FOOD); GROWING THE WORMS; AFTER THE CULTURE HAS GROWN; PREPARING EGGS TO START SYNCHRONIZED LIQUID CULTURES. - [Read Liquid Culture of Worms Protocol]

Microinjection of DNA into Pronucleus of Fertilized Eggs Protocol - http://www.bcm.edu/dtmc/?pmid=7785

Category: Microinjection Protocols: Microinjection of DNA Protocols

Protocol for microinjection of DNA into pronucleus of fertilized eggs. - [Read Microinjection of DNA into Pronucleus of Fertilized Eggs Protocol]

Natural and Artificial Fertilization of Xenopus Eggs Protocol - http://www.hhmi.ucla.edu/derobertis/protocol_page/Pdfs/Natural%20and%20Artificial%20Fertilization%20of%20Frog%20Eggs.pdf

Category: Xenopus Protocols: Xenopus Injection Protocols

Protocol for natural and artificial fertilization of Xenopus eggs. - [Read Natural and Artificial Fertilization of Xenopus Eggs Protocol]

Protocol Spindle Assembly In Vitro - http://www.bio.com/protocolstools/protocol.jhtml?id=p1051

Category: Cell Biology and Cell Culture: In Vitro Reconstitution Protocols

This protocol describes an in vitro reaction to assay mitotic spindle assembly. The assay uses Cytostatic Factor extract made from Xenopus eggs, fluorescently-labelled tubulin, and prepared sperm nuclei. Spindle assembly is monitored by immunofluorescence microscopy. - [Read Protocol Spindle Assembly In Vitro]

Xenopus Egg CSF Extract Preparation Protocol - http://www.bio.com/protocolstools/protocol.jhtml?id=p1049

Category: Xenopus Protocols: Xenopus Egg Extracts Protocols

Protocol describes the preparation of Cytostatic Factor (CSF) from frog eggs. CSF can be used in in vitro spindle assembly assays. Includes: Frog injections; Preparation Before Starting the Extract Preparation; Extract Preparation. - [Read Xenopus Egg CSF Extract Preparation Protocol]

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In Vitro Translated Xenopus Mos Kinase Assay Protocol

In Vitro Translated Xenopus Mos Kinase Assay Protocol. In response to progesterone, immature Xenopus oocytes mature to eggs that can be fertilized. The Mos protein kinase is essential for oocyte maturation, most likely due to its ability to activate the MAP kinase cascade. This MAP kinase cascade eventually leads to the activation of Cdc2/cyclin B and entry into M phase. In this protocol, tagged Mos kinase is translated in vitro, immunopurified, and used in a kinase assay.

Histone H1 Kinase Activity Assay

Histone H1 Kinase Activity Assay Protocol. This protocol describes assaying kinase activity of a putative kinase using Histone H1 as the substrate. Histone H1 is the canonical kinase substrate in this type of assay. Phosphorylation of Histone H1 is assessed by SDS-Polyacrylamide gel electrophoresis followed by autoradiography.

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