Automated system for capture and detection of nucleic acids.

abstract：:The conventional method for cloning a DNA fragment is to insert it into a vector and ligate it. Although this method is commonly used, it is labor intensive because the ratio and concentrations of the DNA insert and the vector need optimizing. Even then, the resultant library is often plagued with unwanted plasmids th...

journal_title：BioTechniques

authors： Miyazaki K,Takenouchi M

更新日期：2002-11-01 00:00:00

abstract：:Here we describe a method for the isolation of PCR-ready genomic DNA from various zebrafish tissues that is based on a previously published murine protocol. The DNA solutions are of sufficient quality to allow PCR detection of transgenes from all commonly used zebrafish tissues. In sperm, transgene amplification was s...

journal_title：BioTechniques

authors： Meeker ND,Hutchinson SA,Ho L,Trede NS

更新日期：2007-11-01 00:00:00

abstract：:The resolution of complex protein mixtures by discontinuous buffer SDS-PAGE is accomplished by their concentration into thin bands in the stacking gel, followed by their separation during migration through the resolving gel. Recombinant human interferon-inducible protein-10 (IP-10), a 10-kDa C-X-C chemokine with four ...

journal_title：BioTechniques

authors： Crow MK,Karasavvas N,Sarris AH

更新日期：2001-02-01 00:00:00

abstract：:Here, we present a rapid and reproducible procedure based on square-wave pulse electroporation that allows efficient penetration of synthetic oligonucleotides into intact yeast cells. This procedure was successfully used to modify the yeast genome with small amounts of oligonucleotide. ...

journal_title：BioTechniques

authors： Barre FX,Mir LM,Lécluse Y,Harel-Bellan A

更新日期：1998-08-01 00:00:00

abstract：:We describe a method using an inexpensive craft glue to routinely isolate specific areas of tissue as small as 1 mm2 from paraffin sections. The tissue may be digested to release nucleic acid suitable for PCR or reverse transcription PCR. The use of this procedure obviates the requirement for manual microdissection or...

journal_title：BioTechniques

authors： Turbett GR,Barnett TC,Dillon EK,Sellner LN

更新日期：1996-05-01 00:00:00

abstract：:Here we introduce a modified antibody staining method that uses up to 80% less antibody for flow cytometry. We demonstrate this method for the detection of antigens expressed at high, moderate, or low levels in mouse and rat lymphocytes as well as mouse mammary epithelial cells. We obtained reproducibly accurate resul...

journal_title：BioTechniques

authors： Sharma D,Eichelberg MR,Haag JD,Meilahn AL,Muelbl MJ,Schell K,Smits BM,Gould MN

更新日期：2012-07-01 00:00:00

abstract：:Under-agarose chemotaxis has been used previously to assess the ability of neutrophils to respond to gradients of chemoattractant. We have adapted this assay to the chemotactic movement of Dictyostelium amoebae in response to folic acid. Troughs are used instead of wells to increase the area along which the cells can ...

journal_title：BioTechniques

authors： Laevsky G,Knecht DA

更新日期：2001-11-01 00:00:00

abstract：:Sarah Webb explores how RNA sequencing innovations help scientists tackle the complexity of the transcriptome. ...

journal_title：BioTechniques

authors： Webb S

更新日期：2018-02-01 00:00:00

abstract：:Recombinant vaccinia viruses (rVACVs) are promising antigen-delivery systems for vaccine development that are also useful as research tools. Two common methods for selection during construction of rVACV clones are (i) co-insertion of drug resistance or reporter protein genes, which requires the use of additional selec...

journal_title：BioTechniques

authors： Liu L,Cooper T,Eldi P,Garcia-Valtanen P,Diener KR,Howley PM,Hayball JD

更新日期：2017-04-01 00:00:00

abstract：:Metabolic and electrical coupling through gap junction channels is implicated in cell differentiation, tissue homeostasis, and electrotonic propagation of signals in excitable tissues. The characterization of gating properties of these channels requires electrophysiological recordings at both single- and multiple-chan...

journal_title：BioTechniques

authors： Zhong G,Mantel PL,Jiang X,Jarry-Guichard T,Gros D,Labarrere C,Moreno AP

更新日期：2003-05-01 00:00:00

abstract：:We have developed an improved subtractive hybridization method that provides a fast, simple and reliable isolation of desired different sequences from two compared DNA libraries, one of which contains all unwanted homologues (subtracter) and another contains certain desired heterologues (tester). The DNA library can b...

journal_title：BioTechniques

authors： Ying SY,Lin S

更新日期：1999-05-01 00:00:00

abstract：:Recombinant retroviruses are a common vehicle to deliver an exogenous gene to a target cell, which makes them a useful tool in the field of gene therapy. A major drawback to using recombinant retroviruses is the low titer achieved, resulting in a limited number of target cells infected and subsequently poor expression...

journal_title：BioTechniques

authors： Tobias CA,Kim D,Fischer I

更新日期：2000-10-01 00:00:00

abstract：:Genomic analysis of circulating, cell-free DNA (cfDNA) is being used extensively for molecular diagnostics. Many approaches rely on the construction of cfDNA genomic libraries, targeted retrieval of specific genomic regions and analysis by next-generation DNA sequencing. Several steps during sample preparation require...

journal_title：BioTechniques

authors： Raymond CK

更新日期：2019-10-01 00:00:00

abstract：:S-100B is used in melanoma follow-up. This serum biomarker is also present in adipocytes; therefore, subcutaneous adipocytes trapped in the needle before performing a venipuncture could contaminate the serum. The aim was to study the influence of adipocyte contamination on blood samples used for S-100B analysis, possi...

journal_title：BioTechniques

authors： Damude S,Muller Kobold AC,Bastiaannet E,Kruijff S,Hoekstra HJ,Wevers KP

更新日期：2020-11-01 00:00:00

abstract：:The concentration of proteins in cells is an important parameter that determines how a protein will interact with other proteins or pharmacological agents. Recent developments in Western blotting techniques have now made this a method of choice to measure protein concentration in complex solutions such as total cell e...

journal_title：BioTechniques

authors： Fradelizi J,Friederich E,Beckerle MC,Golsteyn RM

更新日期：1999-03-01 00:00:00

abstract：:We systematically evaluated the performance and reliability of several widely used, commercially available actin-filament probes in a highly motile breast adenocarcinoma cell line to optimize the visualization of F-actin-rich dynamic lamellipodia. We evaluated four Phalloidin-fluorophores, two anti-actin antibodies, a...

journal_title：BioTechniques

authors： DesMarais V,Eddy RJ,Sharma VP,Stone O,Condeelis JS

更新日期：2019-03-01 00:00:00

abstract：:For a feasible and cost-effective impedance measurement of cellular alterations in real-time, we combined commercially available microelectrode arrays (MEAs), consisting of 60 microelectrodes, with a conventional impedance analyzer. For proof of principle, a breast carcinoma cell line (MCF-7) was cultured on MEAs, and...

journal_title：BioTechniques

authors： Rothermel A,Nieber M,Müller J,Wolf P,Schmidt M,Robitzki AA

更新日期：2006-10-01 00:00:00

abstract：:When PCR is carried out in a polyacrylamide gel, each target molecule forms a molecular colony that comprises many copies of the original template. By counting the number of colonies, one can directly determine the target titer, with 100% of the DNA molecules and approximately 15% of the RNA molecules being detected. ...

journal_title：BioTechniques

authors： Chetverina HV,Samatov TR,Ugarov VI,Chetverin AB

更新日期：2002-07-01 00:00:00

abstract：:We have utilized 5-bromo-2'deoxyuridine (BrdU) substituted DNA as a probe for a number of applications including, principally, for chromosome painting by fluorescence in situ hybridization (FISH) but also for DNA end-labeling to detect apoptotic cell death and for filter hybridization. Br-dUTP was used as a substitute...

journal_title：BioTechniques

authors： Mühlmann-Díaz MC,Dullea RG,Bedford JS

更新日期：1996-07-01 00:00:00

abstract：:The use of reverse transcription (RT) PCR for relative quantitation of gene transcripts relies on the reproducibility of the individual RT, PCR and product measurement steps. Semi-competitive RT-PCR (RT-cPCR) uses an internal competitor template in the PCR step to improve quantitation. We have surveyed the reproducibi...

journal_title：BioTechniques

authors： Hall LL,Bicknell GR,Primrose L,Pringle JH,Shaw JA,Furness PN

更新日期：1998-04-01 00:00:00

abstract：:Hereditary hemochromatosis (HHC) represents an autosomal recessive disease in which increased iron absorption causes iron overload and irreversible tissue damage. The recently detected association between two point mutations in the HFE gene on chromosome 6p and HHC has made it possible to screen for the disease before...

journal_title：BioTechniques

authors： Bosserhoff AK,Seegers S,Hellerbrand C,Schölmerich J,Büttner R

更新日期：1999-06-01 00:00:00

abstract：:A dot immunoblotting technique has been developed to estimate the relative expression levels of tagged recombinant human proteins in mammalian cell culture media. Variations in sample denaturation, blocking agents and membrane composition and treatment were used to optimize the signal-to-noise ratio of the defined pro...

journal_title：BioTechniques

authors： Ellsworth JL,Hamacher N,Bagwell N,West JW

更新日期：2000-05-01 00:00:00

abstract：:Fluorescent live imaging of cells and embryos at subcellular resolution poses significant challenges for biologists due to morbidity and mortality ensuing from phototoxicity. Here we report the use of a spinning-disk confocal microscope to image mouse and bovine preimplantation embryos without impairing their developm...

journal_title：BioTechniques

authors： Ross PJ,Perez GI,Ko T,Yoo MS,Cibelli JB

更新日期：2006-12-01 00:00:00

abstract：:Expanding applications of cDNA microarrays such as fine needle aspiration biopsy and laser capture microdissection necessitate the ability to perform arrays with minute starting amounts of RNA. While methods for amplifying RNA have been advocated, the fidelity of array results using amplified material has not been ful...

journal_title：BioTechniques

authors： Feldman AL,Costouros NG,Wang E,Qian M,Marincola FM,Alexander HR,Libutti SK

更新日期：2002-10-01 00:00:00

abstract：:Presepsin is a 13-kDa N-terminal glycoprotein of CD14. Previously, agitation-induced increases in presepsin levels have been reported; however, the mechanism remains poorly understood. In this study, we aimed to reveal the mechanism of presepsin increase. The agitated plasma or serum was separated using gel exclusion ...

journal_title：BioTechniques

authors： Morino G,Takahashi G,Kan S,Inoue Y,Sato K,Shirakawa K

更新日期：2021-01-29 00:00:00

abstract：:Another method has been developed to attach synthetic peptides to solid supports for use in enzyme immunoassays. The method is based on passively adsorbing a synthetic peptide to a solid-phase support, then further attaching more of the same peptide by means of cross-linking to the previously adsorbed peptide. This me...

journal_title：BioTechniques

authors： Leahy DC,Shah DO,Todd JA

更新日期：1992-11-01 00:00:00

abstract：:A 200-year-old observation might provide a new way to eliminate tumors by infecting cancer patients with bacteria. Kristie Nybo explores a new approach that could transform cancer treatment. ...

journal_title：BioTechniques

authors： Nybo K

更新日期：2018-01-01 00:00:00

abstract：:There is growing consensus on the potential use of pharmacogenetics in clinical practice, and hopes have been expressed for application to the improvement of global health. However, two major challenges may lead to widening the "biotechnological gap" between the developing and the industrial world;first the unaffordab...

journal_title：BioTechniques

authors： Dorado P,Cáceres MC,Pozo-Guisado E,Wong ML,Licinio J,Llerena A

更新日期：2005-10-01 00:00:00

abstract：:Screening methods for chemotherapeutic agents usually rely on the cytotoxic properties of the drugs. However, agents that inhibit invasion may have more efficacy and cause fewer side effects. Various cellular invasion assays have been used to evaluate these types of compounds, including the modified Boyden chamber, mo...

journal_title：BioTechniques

authors： Sasaki CY,Passaniti A

更新日期：1998-06-01 00:00:00

abstract：:We report a robust method for studying en masse changes in translation using cDNA arrays. The relative distribution of messenger RNAs (mRNAs) along polysome gradients was monitored by performing cDNA array analysis of each gradient fraction and quantifying the mRNA translational status by regression analysis. Using th...

journal_title：BioTechniques

authors： Mazan-Mamczarz K,Kawai T,Martindale JL,Gorospe M

更新日期：2005-07-01 00:00:00