Showing papers in "Methods in 2001"

TL;DR: The 2-Delta Delta C(T) method as mentioned in this paper was proposed to analyze the relative changes in gene expression from real-time quantitative PCR experiments, and it has been shown to be useful in the analysis of realtime, quantitative PCR data.

TL;DR: Benefits of real-time PCR include enhanced sensitivity, high throughput, use of a closed-tube system, reduced variation, the ability to simultaneously multiplex reactions, and lack of post-PCR manipulations.

TL;DR: The TAP method is developed as a tool that allows rapid purification under native conditions of complexes, even when expressed at their natural level, and is a very useful procedure for protein purification and proteome exploration.

TL;DR: The real-time RT-PCR technique is very accurate and sensitive, allows a high throughput, and can be performed on very small samples; therefore it is the method of choice for quantification of cytokine profiles in immune cells or inflamed tissues.

TL;DR: Methods for the interpretation of the MEG signals include the popular point current dipole, minimum norm methods, spatial filtering, beamformers, MUSIC, and Bayesian techniques, which are illustrated in examples of interictal epileptic spiking and voluntary hand-motor activity.

TL;DR: Practical considerations for its implementation including experimental apparatus, fluorescent probe selection, surface immobilization, single-molecule FRET analysis schemes, and interpretation are discussed.

TL;DR: A FRET microscopy method that can be used to determine whether proteins that are colocalized at the level of light microscopy interact with one another, implemented using digital microscopy systems such as a confocal microscope or a wide-field fluorescence microscope coupled to a charge-coupled camera.

TL;DR: Results indicate that cell-specific expression of Cre-ER(T) in transgenic mice can be used for efficient tamoxifen-dependent Cre-mediated recombination at loci containing loxP sites, to generate site-specific somatic mutations in a spatiotemporally controlled manner.

TL;DR: This review focuses on the extraction of nucleic acids from fixed and embedded biopsies using both conventional approaches and laser-assisted microdissection and the subsequent application of real-time PCR methods for quantification of mRNA transcripts, gene copy number, and the methylation status.

TL;DR: The fundamental requirements for the creation, isolation, and utilization of TAT-fusion proteins to affect mammalian cells are described, relating to cell cycle progression, apoptosis, and cellular architecture.

TL;DR: Results suggest that genes identified by DNA arrays with a two to fourfold difference in expression cannot be accepted as true or false without validation, and the real-time RT-PCR approach is well suited for validation of differential expression since it is quantitative and rapid and requires 1000-fold less RNA than conventional assays.

TL;DR: The ability to multiplex PCR by probe color and melting temperature (T(m)) greatly expands the power of real-time analysis and creates a "virtual" two-dimensional multiplexing array without the need for an immobilized matrix of probes.

TL;DR: The protocols for a semiautomated, high-throughput, Gal4-based yeast two-hybrid system to generate functional annotations for predicted proteins that so far have remained uncharacterized are described.

TL;DR: This article reviews techniques that allow us to visualize these protein interactions as they occur in living cells and describes the use of FRET microscopy to examine where transcription factors are assembled in the nucleus.

TL;DR: Real-time PCR amplification techniques are currently used to determine the viral load in clinical samples for an increasing number of targets and issues related to standardization and quality control must be resolved to enable routine implementation of these technologies in molecular diagnostics.

TL;DR: Practical methods that are now available to allow the determination of the complexity and scaling relationships of anatomical and physiological patterns, including fractal dimensionality are reviewed.

TL;DR: This work discusses and presents how to design and use molecular beacons to achieve reliable SNP genotyping and allele discrimination in real-time PCR, and provides a new means of analyzing data outputs from such real- time PCR assays that compensates for differences between sample condition, assay conditions, variations in fluorescent signal, and amplification efficiency.

TL;DR: A series of methodological developments aimed at enhancing the power of the methods needed to record simultaneously from populations of neurons over broad regions of the brain in awake, behaving animals are described.

TL;DR: MethyLight is a sensitive, fluorescence-based real-time PCR technique that is capable of quantitating DNA methylation at a particular locus by using DNA oligonucleotides that anneal differentially to bisulfite-converted DNA according to the methylation status in the original genomic DNA.

TL;DR: Methods developed for performing in situ hybridization with immunocytochemistry to localize tissue-specific antigens in zebrafish are described in this article.

TL;DR: A method for construction of high-complexity random peptide libraries is presented and the key steps are highly efficient binary ligation under conditions where the vector is relatively dilute, with only a modest molar excess of insert, followed by efficient electrotransformation into Escherichia coli.

TL;DR: How to use self-splicing inteins in protein semisynthesis and backbone cyclization is described, which involves the precise cleavage and formation of peptide bonds.

TL;DR: Gene transfer by in ovo electroporation has been applied to the study of developmental biology, especially to central nervous system (CNS) development, and misexpression of Pax-5 is shown as an example.

TL;DR: Electroporation of plasmid DNA following whole-embryo culture (WEC) has several advantages including rapid gene expression, minimal laboratory work, precisely targeted regions, and no risk for human beings.

TL;DR: This work describes its preferred method of real-time functional MRI and some of the early results it has obtained with its use, as well as describing here the methods used for statistical processing of these image time series.

TL;DR: A new approach for RNA synthesis that is now as reliable and efficient as DNA synthesis methods is described in this report, which has enabled the routine high-quality synthesis of RNA oligonucleotides up to 50 bases in length regardless of sequence or secondary structure.

TL;DR: The basic approach to analyzing time courses is described, together with some examples of the essential computer code to implement these analyses, and a general method that can be applied to both steady state and non-steady-state systems is recommended.

TL;DR: The isolation of associated proteins from a complex cell-derived lysate by using an epitope-directed antibody was reported, and the protein pICLn engineered to carry an epitopes tag was purified from cultured human embryonic kidney cells, and found to associate with a variety of proteins including the spliceosomal proteins smE and smG.

TL;DR: A number of two-hybrid-based approaches are summarized, and the use of the forward yeast two- Hybrid system in a screen to identify novel interacting partners for a protein of interest is detailed.