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Coliforms
Time of Update: 2021-02-15
The coliform group of indicator organisms includes some members of this family that are capable of fermenting lactose with the production of acid and gas within 48 h at 35�C. The family Enterobacteri
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Detection of Bacterial Protein Toxins by Solid Phase Magnetic Immunocapture and Mass Spectrometry
Time of Update: 2021-02-15
We have shown that staphylococcal enterotoxins, for example, can be efficiently enriched by means of magnetic immunocapture using antibody functionalized paramagnetic beads.
The method is applicable to any bacterial toxin to which an antibody is available.
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Identification and Verification of microRNAs by High-Throughput Sequencing
Time of Update: 2021-02-15
High-throughput sequencing methods have become invaluable for detection and analysis of small RNAs. The results are millions of sequences that need to be carefully analyzed by computational methods a
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Experimental Manipulation of the Microbial Functional Amyloid Called Curli
Time of Update: 2021-02-15
Here we describe experimental methods to study the biogenesis and assembly of curli by exploiting their amyloid properties.
We also present methods to analyze curli-mediated biofilm formation.
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High Isoprenoid Flux Escherichia coli as a Host for Carotenoids Production
Time of Update: 2021-02-15
coli was engineered for high production of carotenoids.
The resulting high isoprenoid fluxE.
In this study, the high isoprenoid fluxE.
coli was transformed with a plasmid carrying the β-carotene biosynthetic genes fromPantoea stewartii for β-carotene production .
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Cell Survival and Apoptosis Assays for E1B 19K and Bcl-2 Family Members
Time of Update: 2021-02-15
Apoptosis is regulated by a number of genes that promote or suppress cell death.
Here, we describe highly reproducible, simple methods that we have developed to assay the activities of adenovirus E1B 19 kDa, Bcl-2, and various pro-apoptotic interacting partners.
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Small scale DNA preps for Neurospora crassa.
Time of Update: 2021-02-15
5. Add 300 microliters of 7.5 M ammonium acetate, invert each tube to mix the contents and incubate on ice for 10 min.
7. Add 500 microliters of isopropanol to each sample, invert the tubes to mix the contents and incubate on ice for 10 min.
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Isolation of Mutants and Construction of Intersexual Heterokaryons of Blakeslea trispora
Time of Update: 2021-02-15
Two genetic techniques have been used to increase carotene accumulation: the isolation of mutants and the formation and segregation of heterokaryons.
Because all life stages are multinucleated, recessive mutants are isolated after exposure toN -methyl- N ′-nitro- N -nitrosoguanidine, a strong mutagen and inactivator of nuclei.
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BAC Rescue: A Tool for Functional Analysis of the Mouse Genome
Time of Update: 2021-02-15
Such mice are generated by introducing a gene of interest into the genome and ectopically expressing it using a heterologous promoter.
Bacterial artificial chromosomes (BACs) contain large segments of genomic DNA (>300 kb) and thus, most likely, contain all thecis -regulatory elements of a gene ( 4 ).
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Using the TIGR Assembler in Shotgun Sequencing Projects
Time of Update: 2021-02-15
This extensive experience led to a sequence assembler that produces few misassemblies ( 2 , 3 ) and has been used successfully in whole-genome shotgun sequencing of prokaryotic and eukaryotic organisms, bacterial artificial chromosome-based sequencing of eukaryotic organisms, and expressed sequence tag assembly.
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Campylobacter: Isolation, Identification, and Preservation
Time of Update: 2021-02-15
The methods described herein apply a microaerophilic environment complemented with supplements such as blood, charcoal, and ferrous sulfate, sodium metabisulfate, and sodium pyruvate (FBP), which are thought to act by quenching toxic oxygen derivatives that develop over time in the media.
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PCR-Detection of Hemophilus paragallinarum, Hemophilus somnus, Mannheimia (Pasteurella) hemolytica, Mannheimia spp., Pasteurella trehalosi, and Pasteurella multocida
Time of Update: 2021-02-15
Out of the almost 100 species or species-like taxa that might be isolated from mammals, reptiles, and birds, onlyPasteurella multocida, Actinobacillus pleuropneumoniae , andHemophilus paragallinarum are regarded as major pathogens ( 1 , 2 ) whileA.
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Real-Time PCR Assay for the Diagnosis of Pneumocystis jirovecii Pneumonia
Time of Update: 2021-02-15
jirovecii cannot be cultured, specific identification of it depends on examining respiratory specimens.
jirovecii not detectable by routine histochemical staining.
We have shown that the direct immunofluorescence assay can be replaced by a real-time PCR assay given its feasibility, sensitivity, and specificity, for the detection ofP.
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An SDS-PAGE-Based Enzyme Activity Assay for the Detection and Identification of Histone Acetyltransferases
Time of Update: 2021-02-15
Posttranslational acetylation of the core histone amino-terminal tails correlates with both chromatin assembly and gene expression. This energy-intensive and reversible process is mediated by the opp
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Interaction Between Lactic Acid Bacteria and Gastrointestinal Nematodes of Caprine Origin
Time of Update: 2021-02-15
To compare the level of parasitism with gastrointestinal nematodes in sheep and goats, several studies have been conducted ( 1 – 3 ).
They have generally shown that goats were more infected than sheep, as they exhibited higher worm burdens and egg excretion.
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Dendritic Cell Generation from Highly Purified CD14+Monocytes
Time of Update: 2021-02-15
Dendritic cells (DC) play a pivotal role in the function of the immune system, for they are the primary antigen-presenting cells (APC) in the activation of naive T-lymphocyte responses ( 1 ).
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Identification of Mycoplasmas by Dot Immunobinding on Membrane Filtration (MF Dot)
Time of Update: 2021-02-15
The definitive identification of mycoplasmas is usually based on serological procedures, including growth inhibition ( 1 , 2 ), metabolic inhibition ( 3 , 4 ), immunofluorescence ( 5 – 7 ), and immunobinding assays ( 8 – 14 ).
All these assay systems are based on the detection of mycoplasma surface antigens, which are believed to be highly specific.
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Analysis of Glycoproteins of Viruses in the Family Bunyaviridae
Time of Update: 2021-02-15
The glycoproteins are modified by N-linked glycosylation and accumulate in the Golgi complex where virions mature and bud.
In this chapter, we describe the methods that have been used in our laboratory for the study of the glycoproteins of Bunyamwera virus, the prototype of the family.
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Detection of Tetracycline Resistance Genes by PCR Methods
Time of Update: 2021-02-15
Therefore, development of genotyping methods for detection of antibiotic resistance genes is highly desirable for fast, accurate, and sensitive detection of antibiotic resistance genes in a broad range of pathogenic and commensal bacteria in both clinical and environmental samples.
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Ultrasensitive ELISA for Measurement of Human Cytokine Responses in Primary Culture
Time of Update: 2021-02-15
In this chapter, we review the major factors to be considered in the development and application of ultrasensitive ELISAs to the analysis of human immune responses.
