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A Rapid Method for Generating Gene Deletions in Aspergillus fumigatus Protocol - http://www.aspergillus.org.uk/indexhome.htm?secure/laboratory_protocols/./genedel.html~main

Category: Fungi Protocols: Aspergillus Protocols

The technique makes use of an Escherichia coli strain expressing the redΑßΓ operon under the control of an inducible promoter. This enables the strain to carry out homologous recombination with only 50-60 bp of homologous sequence. The procedure does not require any DNA ligation and is very rapid. It allows a single gene or region on a cosmid to be replaced by a bi-functional selectable marker (having both an E. coli and an A. fumigatus marker). - [Read A Rapid Method for Generating Gene Deletions in Aspergillus fumigatus Protocol]

Bioassay for Determining Itraconazole Levels in Blood - http://www.aspergillus.org.uk/indexhome.htm?secure/laboratory_protocols/./itrabio.html~main

Category: Fungi Protocols

Serum concentrations of itraconazole should be measured in patients receiving this drug to ensure that therapeutic concentrations are being achieved. This is necessary as drug absorption can be variable, and levels may be lowered by interactions with other drugs. The assay will give an indication of whether suitable blood levels have been achieved. - [Read Bioassay for Determining Itraconazole Levels in Blood]

Detection of Fungi by Fluorescence Microscopy Using Fluorescent Brighteners - http://www.aspergillus.org.uk/indexhome.htm?secure/laboratory_protocols/./flbr.html~main

Category: Microscopy Protocols: Fluorescence Microscopy Protocols

Certain fluorescent dyes such as Blankophor have a high affinity for the b -glycosidically linked polysaccharides such as glucan and chitin, which are main the constituents of the fungal cell wall. Therefore, these fluorescent dyes can be used for screening clinical samples for the presence of fungal elements. This procedure can be performed using the following specimens: Nail, Skin, Bronchial alveolar lavage fluid, Sputum and Biopsies. - [Read Detection of Fungi by Fluorescence Microscopy Using Fluorescent Brighteners]

Detection of Fungi by Fluorescence Microscopy Using Fluorescent Brighteners Protocol - http://www.aspergillus.org.uk/indexhome.htm?secure/laboratory_protocols/./flbr.html~main

Category: Fungi Protocols

Protocol for the detection of fungi by fluorescence microscopy using fluorescent brighteners. - [Read Detection of Fungi by Fluorescence Microscopy Using Fluorescent Brighteners Protocol]

Long-PCR Reagents and Guidelines - http://wheat.pw.usda.gov/~lazo/methods/lazo/longpcr.html

Category: PCR Protocols: Quantitative PCR Protocols

General guidelines for long-PCR conditions and enzyme mixtures. Efficient long-PCR results from the use of two polymerases: a non-proofreading polymerase is the main polymerase in the reaction, and a proofreading polymerase (3' to 5' exo) is present at a lower concentration. Includes: For PCR with low-complexity templates (e.g., plasmid and cosmid inserts); For PCR with moderate-complexity templates (e.g., bacterial genomic DNA); For PCR with high-complexity templates (e.g., human genomic DNA). - [Read Long-PCR Reagents and Guidelines]

Method for Assaying Gliotoxin Production in Aspergillus fumigatus Protocol - http://www.aspergillus.org.uk/indexhome.htm?secure/laboratory_protocols/./gliotoxin1.html~main

Category: Fungi Protocols: Aspergillus Protocols

Gliotoxin is a metabolite of Aspergillus fumigatus that exhibits immunosuppressive activity against certain cells of the immune system. Secretion of gliotoxin during infection has been suggested as being a factor in the pathogenesis of aspergillosis. Gliotoxin secretion can be assayed in a number of ways by thin layer chromatography (TLC) high performance liquid chromatography (HPLC) or bioassay using the effect of gliotoxin on human cells1. - [Read Method for Assaying Gliotoxin Production in Aspergillus fumigatus Protocol]

Nucleic Acid Hybridization General Aspects - http://www.roche-applied-science.com/PROD_INF/MANUALS/InSitu/pdf/ISH_33-37.pdf

Category: Cytogenetics Protocols: In Situ Hybridization Protocols

Discusses the effects of various components of the hybridization solution on the rate of renaturation and thermal stability of DNA hybrids free in solution. Includes: The main parameters that influence hybridization; Additional hybridization variables; Competition in situ hybridization; Oligonucleotide hybridization; Standard in situ hybridization conditions. - [Read Nucleic Acid Hybridization General Aspects]

Phagocytosis Assay Protocol - http://www.aspergillus.org.uk/indexhome.htm?secure/laboratory_protocols/./protocol5.html~main

Category: Fungi Protocols

Protocol for phagocytosis assay. Protocol describes how to assess the efficiency of phagocytic cells to engulf conidia of a given fungus. - [Read Phagocytosis Assay Protocol]

Preparation of Embryonic Stem (ES) Cells for Injection Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4416

Category: Stem Cell Protocols: Stem Cell Injection Protocols

Injection of ES cells into the cavity of blastocysts is one of the main methods for generating chimeras. This protocol describes the preparation of ES cells for injection. - [Read Preparation of Embryonic Stem (ES) Cells for Injection Protocol]

Preparation of Solid Tissue for Aspergillus Galactomannan Antigen Detection by Platelia Protocol - http://www.aspergillus.org.uk/indexhome.htm?secure/laboratory_protocols/./galactoman.html~main

Category: Fungi Protocols

Protocol for the preparation of solid tissue for Aspergillus galactomannan antigen detection by Platelia (Biorad). Technique was designed for use on human serum. However, it may also be possible to perform this method on solid tissues and organic solutions. Viscous solution and tissue specimens need to be pre-treated to achieve the extraction of the Aspergillus antigen and to get a homogeneous sample in solution. - [Read Preparation of Solid Tissue for Aspergillus Galactomannan Antigen Detection by Platelia Protocol]

Protocol for L-selectin-PNAd Interactions under Flow Conditions. - http://www.cbrinstitute.org/labs/springer/protocols/azucena_flow_chamber.html

Category: Cell Biology and Cell Culture

This protocol focuses on the interactions between L-selectin expressed on neutrophils and PNAd coated onto the plastic surface. The main purpose of the flow chamber assay is to visualize and measure interactions between flowing cells expressing a given adhesion molecule on their surface, and their receptor, either directly coated on the flow chamber lower wall or expressed on a cell monolayer. - [Read Protocol for L-selectin-PNAd Interactions under Flow Conditions.]

Protocol: Bioassay for Determining Voriconazole Levels in Blood - http://www.aspergillus.org.uk/indexhome.htm?secure/laboratory_protocols/./BIOASSAY.html~main

Category: Fungi Protocols

Serum concentrations of voriconazole should be measured in patients receiving this drug to ensure that therapeutic levels are being achieved. The assay will give an indication of whether suitable blood levels have been achieved. - [Read Protocol: Bioassay for Determining Voriconazole Levels in Blood]

Protoplast Preparation for A nidulans Protocol - http://www.aspergillus.org.uk/indexhome.htm?secure/laboratory_protocols/./protoplastRL.html~main

Category: Fungi Protocols: Aspergillus Protocols

Protocol for protoplast preparation for A nidulans. - [Read Protoplast Preparation for A nidulans Protocol]

Purification of Monocytes Protocol - http://www.aspergillus.org.uk/indexhome.htm?secure/laboratory_protocols/./protocols.html~main

Category: Immunology: Mononuclear Cell Isolation Protocols

Protocol for the purification of monocytes. - [Read Purification of Monocytes Protocol]

Purification of Neutrophils Protocol - http://www.aspergillus.org.uk/indexhome.htm?secure/laboratory_protocols/./protocol1.html~main

Category: Immunology: Mononuclear Cell Isolation Protocols

Protocol for purification of neutrophils. - [Read Purification of Neutrophils Protocol]

Quality Control Considerations for Cell Culture - http://www.sigmaaldrich.com/Area_of_Interest/Life_Science/Cell_Culture/Key_Resources/ECACC_Handbook/Cell_Culture_Techniques_9.html#Quality%2520Control

Category: Cell Culture Protocols: Cell Culture Contamination Prevention and Removal Protocols

Quality is important in all aspects of tissue culture since the quality of materials used i.e. media and other reagents) will affect the quality of the cultures and products derived from them. The main areas of quality control that are of concern for tissue culture are: The quality of the reagents and materials; The provenance and integrity of the cell lines; The avoidance of microbial contamination. - [Read Quality Control Considerations for Cell Culture]

Random Amplification of Polymorphic DNA (RAPD) Typing and Fingerprinting Protocol - http://www.aspergillus.org.uk/indexhome.htm?secure/laboratory_protocols/./rapds.html~main

Category: Fungi Protocols: Aspergillus Protocols

RAPD is a procedure for typing and fingerprinting isolates of a species. It can be used for epidemiological studies, such as investigations into hospital outbreaks and as a laboratory aid to keep track of cultures and to verify that mutants generated in the laboratory are genetically identical to the parental strain. In our hands, the use of one primer, R108, is sufficiently discriminatory to distinguish between the isolates of different strains. - [Read Random Amplification of Polymorphic DNA (RAPD) Typing and Fingerprinting Protocol]

Sampling of Aspergillus spp Spores in Air Protocol - http://www.aspergillus.org.uk/indexhome.htm?secure/laboratory_protocols/./airprotocol.html~main

Category: Fungi Protocols: Aspergillus Protocols

Protocol allows the isolation and enumerate Aspergillus spp spores in air. Includes: Sampling Procedure; Sampling Location; Selection of Sampling Time; Sampling Steps; Laboratory Procedure; Enumerating the Colony Forming Units. - [Read Sampling of Aspergillus spp Spores in Air Protocol]

TAIL PCR Protocol - http://preuss.bsd.uchicago.edu/protocols/tail.html

Category: DNA Protocols: PCR Polymerase Chain Reaction Protocols

TAIL PCR Protocol. TAIL is a series of reactions that are intended to map where a T-DNA (transfer DNA) has inserted within the genome. The main components of the 3 reactions are the AD (Arbitrary Degenerate) primers, border primers, and DNA from the T-DNA - [Read TAIL PCR Protocol]

TAIL PCR Protocol - http://preuss.bsd.uchicago.edu/protocols/tail.html

Category: PCR Protocols

Articles (1-5 of 5)

Probe Preparation for 22 x 40 mm DNA Microarrays

Probe Preparation for 22 x 40 mm DNA Microarrays Protocol.

Antibodies Phage Expression Protocol Using 96-well Microtiter Plates

A Phage display expression protocol for antibody expression in 96-well microtiter plates.

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.

Thionin Method

Thionin Method and protocol for staining cells.

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.

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