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Generation of BAC Transgenic Mice
Time of Update: 2021-01-26
Transgenic mice provide an important experimental system for studying the effects of gene overexpression and for analyzing the expression of well-characterized “single” genes that contribute to a particular trait.
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DNA Amplification: General Concepts and Methods
Time of Update: 2021-01-26
) are more or less confined to diagnostic applications.
For example, the product of a PCR, often referred to as an amplification, can be readily sequenced for diagnostic, typing, fingerprinting, or molecular epidemiologic reasons.
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Transcript Localization in Dictyostelium discoideum Cells by RNA FISH
Time of Update: 2021-01-26
Biogenesis of ribosomal RNA (rRNA) takes place preliminary in the nucleolus of eukaryotic cells, the site of r DNA transcription.
Several processing steps of rRNA molecules have been implied to take place in the cytoplasm.
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Isolation of scFvs that Inhibit the NS3 Protease of Hepatitis C Virus by a Combination of Phage Display and a Bacterial Genetic Screen
Time of Update: 2021-01-26
The methods can be used more generally to isolate protease-inhibiting cytoplasmic intrabodies able to inhibit proteases or other activities that can be linked with the phenotype ofEscherichia coli .
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Making High Titer Retroviral Producer Cells
Time of Update: 2021-01-26
Retroviral vectors are uniquely suitable for high-efficiency gene transfer to a large target cell population and this (among other things) has led to the exploration of previously impractical strategies for somatic gene therapy ( 1 , 2 ).
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Design and Construction of Synthetic Phage-Displayed Fab Libraries
Time of Update: 2021-01-26
Here we describe detailed protocols for the design and construction of phage-displayed synthetic antibody libraries in which diversity is generated in the complementarity determining regions (CDRs) of the V H of a single humanized bivalent Fab scaffold.
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Quantifying Pseudomonas aeruginosa Quinolones and Examining Their Interactions with Lipids
Time of Update: 2021-01-26
Separation of PQS from extracts by Thin-Layer Chromatography (TLC) is used in combination with the natural fluorescence of the molecule for quantification.
A second separation technique for the PQS precursor HHQ using High-Performance Liquid Chromatography (HPLC) is also described, and this assay exploits the molecule’s characteristic absorbance for quantification.
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Radiation Hybrid Mapping With BAC Ends
Time of Update: 2021-01-26
The availability of the human draft sequence ( 1 ) now offers the tool for sequence-based genetic analyses on a genomewide level.
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Rapid Detection and Simultaneous Strain Differentiation of Mycobacterium tuberculosis Complex Bacteria by Spoligotyping
Time of Update: 2021-01-26
The discovery of polymorphicDNA inMycobacterium tuberculosis has led to several methods to differentiate climcal isolates.
The most widely used method is based on the mobility of the insertion element IS 6110 , which is present in virtually allM tuberculosis isolates, usually in multiple copies.
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Quantifying Plasmid Copy Number to Investigate Plasmid Dosage Effects Associated with Directed Protein Evolution
Time of Update: 2021-01-26
We have observed that when random mutations are not restricted to the coding sequence of the target genes, directed evolution results in a strong positive selection of plasmid origin of replication ( ori ) mutations.
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Interaction Between Respiratory Syncytial Virus and Glycosaminoglycans, Including Heparan Sulfate
Time of Update: 2021-01-26
These GAGs are sulfated to varying extents, lending a negative charge, and are used by a large number of viruses to initiate infection of immortalized cell lines.
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Generation of Recombinant Adenovirus Using the Escherichia coli BJ5183 Recombination System
Time of Update: 2021-01-26
The method described here uses homologous recombination machinery ofEscherichia coli BJ5183 to construct plasmids used in generation of adenoviral vectors.
With this method, no ligation steps are involved in generating the plasmids, and any region of the adenoviral genome can be easily modified.
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Isolation and Quantification of Genomic DNA from Mycobacterium tuberculosis
Time of Update: 2021-01-26
3. Centrifuge the Eppendorf tube for 5 min at room temperature using an aerosol-containing microfuge operating at 9,000 rpm.
7. Next, add 100 ml of 5 M NaCl to the cell suspension and thoroughly mix the contents of the Eppendorf tube using a vortex.
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Detection of Mycoplasma hyopneumoniae from Clinical Samples and Air
Time of Update: 2021-01-26
hyopneumoniae occurs by direct contact or aerosol in chronically infected herds when young susceptible pigs are in contact with older pigs.
Piglets can be infected by gilts, low parity sows, but also by older sows ( 2 ).
However, carrier sows represent the most likely source of transmission, ofM.
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Subfractionation and Analysis of the Cell Envelope (Lipo)polysaccharides of Mycobacterium tuberculosis
Time of Update: 2021-01-26
Much of what is known about their biosynthesis was derived from the phenotypic analysis of knockout or conditional knockout mutants of mycobacteria generated by random or specific insertional mutagenesis.
tuberculosis cells and investigate major quantitative and qualitative changes in their cell envelope (lipo)polysaccharides.
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Method for the Quantification of Aquatic Primary Production and Net Ecosystem Metabolism Using In Situ Dissolved Oxygen Sensors
Time of Update: 2021-01-26
We present an example-based methodology for the “open-water” technique to determine net ecosystem metabolism (or net community metabolism) of coastal and oceanic environments using dissolved oxygen measurements collected with in situ sensors.
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Electroporation of Dictyostelium discoideum
Time of Update: 2021-01-26
The ability to introduce molecules into cells has allowed the analysis of promoters by transient and stable expression of reporter genes, such as luciferase and β-galactosidase ( 2 , 3 ).
Stable transformants have also been created that overexpress normal and altered copies of important structural and regulatory proteins ( 4 – 9 ).
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Clostridium difficile and the Disease It Causes
Time of Update: 2021-01-26
difficile strain NAP1/027 emerging in 2002 in Canada and the USA has now spread to most parts of Europe and virulence factors like high toxin production and sporulation challenge the therapeutic situation and cause great concern among infection control workers.
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Comparison of three testing methods for Helicobacter pylori infection
Time of Update: 2021-01-26
In order to find a simple, inexpensive and easy experimental method, we tried to separate, identify and compare hp in gastric biopsy specimens with urea enzyme tests and smear lenses by candlestick, as reported below.
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Defined Media for H. pylori
Time of Update: 2021-01-26
In a series of studies, Mendz, Hazell, and colleagues found clear evidence for the pentose phosphate pathway ( 4 ), glucokinase activity ( 5 ), and the fermentation of glucose to lactate ( 6 ).
