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Clone Genes From a Phage Library Protocol - http://www.ucsf.edu/micro/faculty/Johnson/protocols/protocol1.html

Category: DNA Protocols: cDNA Library Protocols: cDNA Library Construction Protocols

Protocol for cloning genes from a phage library. Includes: Titer and plate out phage; Lift plaques onto filters and prepare them for screening; Make a probe; Hybridize the probe to the filters; Wash the filters and expose to film; Purify putative plaques; Excise plasmid from the desired phage. - [Read Clone Genes From a Phage Library Protocol]

Collecting Blastocysts Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4360

Category: Mouse Protocols: Mouse Embryology Protocols

Protocol describes a method for collecting blastocysts from pregnant female mice at 3.5 to 4.5 days post coitum (dpc). The blastocysts can then be injected with embryonic stem cells to make chimeras. - [Read Collecting Blastocysts Protocol]

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]

Fission Yeast Media Protocol - http://www-rcf.usc.edu/~forsburg/media.html

Category: Cell Biology and Cell Culture: Cell Culture Growth Media: Yeast Cell Culture Growth Media Protocols

Protocol for fission yeast media. Includes: YES (yeast extract with supplements) for rich complete media; YSO, for modified rich media; EMM (Edinburgh minimal medium) for minimal selective media; stock solutions to make up minimal media; MB, a very stringent minimal for transformation; Sporulation media; ME (malt extract); SPAS; EMMGlut. - [Read Fission Yeast Media Protocol]

Flow Cytometric Determination of Leukocyte Surface Antigens in Whole Blood Protocol - http://pingu.salk.edu/flow/protocols/wholeblood.html

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Flow Cytometry Immunofluorescence Staining Protocols

Protocol for flow cytometric determination of leukocyte surface antigens in whole blood. Includes: Blood collection; Add antibody to the blood; Make measurements. - [Read Flow Cytometric Determination of Leukocyte Surface Antigens in Whole Blood Protocol]

Flow Cytometry of Apoptosis Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E663022BBACCC599F244293812ED95A&objectid=66744E3FA79C848C2D8B85BB84708AC9

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Apoptosis Cytometry Protocols

Common methods applicable to flow cytometry make it possible to: (1) identify and quantify dead or dying cells, (2) reveal a mode of cell death (apoptosis or necrosis), and (3) study mechanisms involved in cell death. Gross changes in cell morphology and chromatin condensation, which occur during apoptosis, can be detected by analysis with laser light beam scattering. - [Read Flow Cytometry of Apoptosis Protocol]

Generation of Bidirectional Sets of Deletion Mutants by Digestion with BAL 31 Nuclease Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3589

Category: Genetics and Genomics: Genotyping Protocols: Mutation Detection Protocols

In this method, the nuclease BAL 31 is used to make uni- or bidirectional deletions in a segment of cloned DNA. BAL 31 is a complex enzyme and tends to digest a population of double-stranded DNA targets in an asynchronous fashion, Deletions created by BAL 31 are therefore far more heterogeneous in size than those created by processive enzymes such as exonuclease III. - [Read Generation of Bidirectional Sets of Deletion Mutants by Digestion with BAL 31 Nuclease Protocol]

How to Make a Cross - http://www.fgsc.net/neurosporaprotocols/How%20to%20make%20a%20cross-2.pdf

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to make a cross. - [Read How to Make a Cross]

How to Make Competent Cells - Calcium Chloride - http://sosnick.uchicago.edu/competent_cells.html

Category: Microbiology: Bacterial Competent Cells Preparation E.Coli

How to Make Competent Cells - Calcium Chloride. Sosnick Lab, Univ. Chicago - [Read How to Make Competent Cells - Calcium Chloride]

How to Make Disposable Spreaders from Long-Nose Pasteur Pipets - http://www.fgsc.net/neurosporaprotocols/How%20to%20make%20disposable%20spreaders%20from%20long-nose%20Pasteur%20pipets%20final.pdf

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to make disposable spreaders from long-nose Pasteur pipets. - [Read How to Make Disposable Spreaders from Long-Nose Pasteur Pipets]

How to Make Heterokaryons in Neurospora crassa - http://www.fgsc.net/neurosporaprotocols/How%20to%20make%20heterokaryons%20in%20Neurospora%20crassa.pdf

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to make heterokaryons in Neurospora crassa. - [Read How to Make Heterokaryons in Neurospora crassa]

How to Make Tapered Platinum-Iridium Needles and Use Them for Isolating Ascospores - http://www.fgsc.net/neurosporaprotocols/How%20to%20make%20tools%20for%20spore%20final.pdf

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to make tapered platinum-iridium needles and use them for isolating ascospores. - [Read How to Make Tapered Platinum-Iridium Needles and Use Them for Isolating Ascospores]

How to Make Very Small Inocula - http://www.fgsc.net/neurosporaprotocols/How%20to%20make%20very%20small%20inocula.pdf

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to make very small inocula. - [Read How to Make Very Small Inocula]

Information to Make Antibodies against Peptides - Pierce - http://www.piercenet.com/y/production/producebottom.html

Category: Peptide Protocols: Antibodies from Peptides Protocols

Great resource for information. on making antibodies against peptides - Pierce - [Read Information to Make Antibodies against Peptides - Pierce]

Making Injection Pipettes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4395

Category: Microinjection Protocols: Microinjection Equipment

This protocol describes the determination of useful settings with the Sutter Puller P-97 to make injection pipettes for microinjection. - [Read Making Injection Pipettes Protocol]

Making Injection Pipettes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4395

Category: Mouse Protocols: Mouse Embryology Protocols

Protocol describes the determination of useful settings with the Sutter Puller P-97 to make injection pipettes for microinjection. - [Read Making Injection Pipettes Protocol]

Mini-Chamber for Regulating Gaseous Environment During Culture - http://www.animal.ufl.edu/hansen/protocols/minigaschamber.htm

Category: Developmental Biology: Embryology Protocols

A simple and inexpensive chamber for regulating gaseous environment of small culture plates, such as those used for culture of preimplantation embryos, can be constructed using disposable media-filtration devises such as Corning’s 115-ml system. The following is a description of how to make such a device. - [Read Mini-Chamber for Regulating Gaseous Environment During Culture]

Monosaccharide Composition Analysis: Alditol Acetates Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4246

Category: Chromatography Protocols: Gas Chromatography Protocols

Protocol describes how to determine the monosaccharide composition of glycans, glycoproteins, or proteoglycans by hydrolyzing the sample to monosaccharides and converting them to alditols, then performing acetylation to make them volatile compounds and analysis by gas chromatography (GC) or gas chromatography coupled with mass spectrometry (GC-MS). - [Read Monosaccharide Composition Analysis: Alditol Acetates Protocol]

Protein Expression and Purification Protocol - http://sosnick.uchicago.edu/Protein_Express&Purify.html

Category: Recombinant Protein Protocols

From plasmid to protein using bacterial expression. Transform appropriate DNA plasmid, Make a starter culture for protein expression, bacteria culture for protein expression. Sosnick Group Chicago. - [Read Protein Expression and Purification Protocol]

RUVKUN Antibody Staining Protocol - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=11

Category: Immunohistochemistry Protocols: Antibody Staining Protocols

Protocol for RUVKUN antibody staining. Includes: Fixation; To make solution; REDUCING DISULFIDES TO -SH; OXIDIZE -SH GROUPS TO -SO3; TO CHECK THAT WORMS ARE PERMEABLE TO MACROMOLECULES; ANTIBODY INCUBATIONS; PERMANENT SPRINGTIME MOUNTING. - [Read RUVKUN Antibody Staining Protocol]

RUVKUN Antibody Staining Protocol - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=11

Category: C. elegans Protocols: C. elegans Staining Protocols

Protocol for RUVKUN Antibody Staining. Includes: Fixation; To make solution; REDUCING DISULFIDES TO -SH; OXIDIZE -SH GROUPS TO -SO3; TO CHECK THAT WORMS ARE PERMEABLE TO MACROMOLECULES; ANTIBODY INCUBATIONS; PERMANENT SPRINGTIME MOUNTING. - [Read RUVKUN Antibody Staining Protocol]

Simple Arabidopsis Transformation Protocol - http://www.arabidopsis.org/info/transformation.jsp

Category: Plant Biology Protocols: Arabidopsis Transformation Protocols

Protocol for a simplified Arabidopsis transformation. Found that the MS salts, hormone, etc. make no difference, that OD of bacteria doesn't make much of a difference, that vacuum doesn't even make much of a difference as long as you have a decent amount of surfactant present. Plant health is still a major factor - healthy fecund plants make a big difference! With this method you should be able to achieve transformation rates above 1%. - [Read Simple Arabidopsis Transformation Protocol]

Studying Mitosis in Cultured Mammalian Cells Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/3/pdb.prot4674

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Cell Cycle Analysis Protocols

Protocol describes methods for maintaining healthy, dividing mammalian cells in culture and during imaging, when mitosis can be examined. Rose chambers are preferable for observation and microinjection of living mitotic cells, but slide/coverslip preparations are easy to make and do not require any special equipment. - [Read Studying Mitosis in Cultured Mammalian Cells Protocol]

Studying Mitosis in Cultured Mammalian Cells Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/3/pdb.prot4674

Category: Cytogenetics Protocols

Protocol describes methods for maintaining healthy, dividing mammalian cells in culture and during imaging, when mitosis can be examined. Rose chambers are preferable for observation and microinjection of living mitotic cells, but slide/coverslip preparations are easy to make and do not require any special equipment. Another inexpensive and easy-to-use alternative is to grow cells in a culture dish with a glass bottom. Such dishes are suitable for microinjection experiments. - [Read Studying Mitosis in Cultured Mammalian Cells Protocol]

Articles (1-10 of 11)

Single Stranded Plasmid DNA Isolation Protocol

A Single Stranded Plasmid DNA Isolation Protocol describing the production and isolation of single-stranded DNA (ssDNA) using bacteriophagemid-containing bacteria and helper phage. Infection of the host cells with helper phage allows for packaging of ssDNA into bacteriophage. The ssDNA can then be isolated from phage particles.

Purifying DNA Fragments from Agarose Gels Alternate Protocol

An alternate protocol for purifying DNA fragments from agarose gels.

Quick DNA Agarose Gel Electrophoresis Protocol.

A quick and simple protocol for the electrophoresis of DNA fragments using agarose gels.

Absorption Spectroscopy and Quantification of Filamentous Phage Protocol

Unlike spherical phage, such as T4 and λ, which have roughly equal weight ratios of protein to DNA, filamentous phage have about six times more protein than DNA; the protein therefore contributes substantially to the absorption spectrum.

DNA Ligation Protocol

The DNA Ligation protocol described here contains the steps required to join together using ligase enzyme both plasmid DNA and insert DNA fragments in order to create a new plasmid. This new ligated plasmid can be transformed after into competent bacteria to produce DNA for mini, midi or maxi-prep isolation.

Crystal Violet Stain

Crystal Violet stain preparation method and protocol.

Thionin Staining

Thionin Staining method.

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.

Picking Transfected ES Cell Clones Protocol

This protocol a protocol on how to generate transfected embryonic stem (ES) cell clones. The previous protocol in this series is the Protocol for Electroporation of ES cells. The next protocol in the series is the Protocol on Disaggregation, Expansion, and Freezing of Transfected ES Clones.

Electrotransformation of BMH 81-17mut S for Isolating Site-Directed dsDNA Mutants

This protocol describes the electroporation of the BMH 81-17 mut S strain that is recommended for tranformation of the site directed mutagenesis of dsDNA (See Protocol on Site-Directed Mutagenesis on Double Stranded DNA). BMH 81-17 mut S are a mismatch repair defective (mut S) Escherichia coli strain. The probability that the two mutations will cosegregate during the first round of DNA replication is increased in this strain.

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