A simple method for isolation of DNA from blood clots suited for use in PCR.

abstract：:A new procedure is described for the generation of high-titer, helper-free retrovirus vectors employing receptor-mediated, adenovirus-augmented transfection into a standard packaging cell line. Viral titers are increased 30-fold to 100-fold in transiently (> 10(5) infectious units per mL) and stable (> 10(7) infectiou...

journal_title：BioTechniques

authors： von Rüden T,Stingl L,Cotten M,Wagner E,Zatloukal K

更新日期：1995-03-01 00:00:00

abstract：:Reducing the scale of biochemical reactions is becoming commonplace. Examples include the screening of large libraries of chemical compounds or gene sequences. These applications demand the ability to transfer sub-microliter volumes of fluid. To this end, we have modified a Hamilton MICROLAB 2200 with high-speed solen...

journal_title：BioTechniques

authors： Hicks JS,Harker BW,Beattie KL,Doktycz MJ

更新日期：2001-04-01 00:00:00

abstract：:New cloning vectors have been developed with features to enhance quick allelic exchange in gram-negative bacteria. The conditionally replicative R6K and transfer origins facilitate conjugation and chromosomal integration into a variety of bacterial species, whereas the sacB gene provides counterselection for allelic e...

journal_title：BioTechniques

authors： Mejia-Santana A,Lloyd CJ,Klose KE

更新日期：2021-01-25 00:00:00

abstract：:Virus uptake is the first rate-limiting step for the successful gene delivery of any virus-based gene therapy. For adenovirus-based gene therapy, the expression levels of the adenovirus receptor--coxsackievirus and adenovirus receptor (CAR)--play an important role in dictating gene delivery. We have observed a wide sp...

journal_title：BioTechniques

authors： Lai YJ,Pong RC,McConnell JD,Hsieh JT

更新日期：2003-07-01 00:00:00

abstract：:The high-order structure of human chromosomes is an important biological question that is still under investigation. Studies have been done on imaging human mitotic chromosomes using mostly 2-D microscopy methods. To image micron-sized human chromosomes in 3-D, we developed a procedure for preparing samples for serial...

journal_title：BioTechniques

authors： Yusuf M,Chen B,Hashimoto T,Estandarte AK,Thompson G,Robinson I

更新日期：2014-12-01 00:00:00

abstract：:At many research institutions, lab space is more valuable than gold. Developers are taking note by designing smaller instruments with enhanced capabilities. Nathan Blow looks inside today's tiny lab. ...

journal_title：BioTechniques

authors： Blow NS Ph D

更新日期：2017-01-01 00:00:00

abstract：:Cytokines are pivotal to a balanced innate or cell-mediated immune response, can be indicative of disease progression and/or resolution, and are being evaluated as therapeutics. There is a need to purify and/or to measure key cytokines rapidly with accuracy, precision, and sensitivity. The current assay technologies, ...

journal_title：BioTechniques

authors： Bai H,Buller RM,Chen N,Boyle MD

更新日期：2002-01-01 00:00:00

abstract：:Loop-mediated isothermal amplification (LAMP) is a rapid and reliable sequence-specific isothermal nucleic acid amplification technique. To date, all reported real-time detection methods for LAMP have been restricted to single targets, limiting the utility of this technique. Here, we adapted standard LAMP primers to c...

journal_title：BioTechniques

authors： Tanner NA,Zhang Y,Evans TC Jr

更新日期：2012-08-01 00:00:00

abstract：:DNA sequences containing tandem dinucleotide repeats represent an abundant source of DNA polymorphism in human and other eukaryotic genomes. Here we describe a novel technique for the identification and characterization of regions of DNA containing these repetitive elements. Using primers designed to recognize tandem ...

journal_title：BioTechniques

authors： Grist SA,Firgaira FA,Morley AA

更新日期：1993-08-01 00:00:00

abstract：:Characterization of the thermodynamics of binding interactions is important in improving our understanding of bimolecular recognition and forms an essential part of the rational drug design process. Isothermal titration calorimetry (ITC) is rapidly becoming established as the method of choice for undertaking such stud...

journal_title：BioTechniques

authors： Holdgate GA

更新日期：2001-07-01 00:00:00

abstract：:We describe a multipurpose eukaryotic expression vector that incorporates the following features: restriction sites for in-frame insertion of cDNAs of interest between sequences encoding the glutathione-S-transferase (GST) and an oligohistidine element, allowing expression of the corresponding fusion proteins; a phosp...

journal_title：BioTechniques

authors： Chatton B,Bahr A,Acker J,Kedinger C

更新日期：1995-01-01 00:00:00

abstract：:In this report we present a rapid and inexpensive PCR-based method to screen recombinant DNA libraries. The efficiency of this method was demonstrated by the isolation of clones of interest from three different libraries using different vector systems. This method is nonradioactive and makes it easier to handle a larg...

journal_title：BioTechniques

authors： Amaravadi L,King MW

更新日期：1994-01-01 00:00:00

abstract：:Human telomerase, a ribonucleoprotein enzyme is known to be associated with immortalized cancer cells but is absent in most normal tissues. Thus, telomerase appears to be an attractive new target for anticancer agents and an important diagnostic marker of human cancers. Here, we describe an improved telomerase assay m...

journal_title：BioTechniques

authors： Sun D,Hurley LH,Von Hoff DD

更新日期：1998-12-01 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：:Selective delivery of genes to ocular tissues in vivo has been a long sought after goal for potential gene therapy of ocular disease. The gene gun was considered for this purpose because of its ability to focally transfer DNA to cells through gold microparticles coated with DNA. Through experimentation, we optimized a...

journal_title：BioTechniques

authors： Tanelian DL,Barry MA,Johnston SA,Le T,Smith G

更新日期：1997-09-01 00:00:00

abstract：:Conformation-sensitive gel electrophoresis (CSGE) has been introduced as the most reliable method for the screening of large and multi-exon genes because of its simplicity, sensitivity and specificity. Based on heteroduplex formation and with the use of mildly denaturing solvents, it allows detection of single-base mu...

journal_title：BioTechniques

authors： Blesa JR,Hernández-Yago J

更新日期：2000-05-01 00:00:00

abstract：:The degree of fluorescence polarization (FP) of a fluorescent molecule is a reflection of its molecular weight (Mr). FP is therefore a useful detection methodfor homogeneous assays in which the starting reagents and products differ significantly in Mr. We have previously shown that FP is a good detection method for th...

journal_title：BioTechniques

authors： Hsu TM,Chen X,Duan S,Miller RD,Kwok PY

更新日期：2001-09-01 00:00:00

abstract：:Haplotypes are useful in population genetics and medicine. Determining haplotypes in the absence of DNA samples from appropriate family members can be difficult and laborious. We have developed a rapid and reproducible method for haplotyping an individual in the absence of relatives. The method utilizes pairs of allel...

journal_title：BioTechniques

authors： Sarkar G,Sommer SS

更新日期：1991-04-01 00:00:00

abstract：:Impurity assays for recombinant protein therapeutics are essential to ensure batch-to-batch consistency and to meet the FDA's criteria for a well-characterized biopharmaceutical. For determination of residual host cell DNA, membrane hybridization assays utilizing radiolabeled DNA probes prepared from the host cell's g...

journal_title：BioTechniques

authors： Ji X,Lee K,DiPaolo B

更新日期：2002-05-01 00:00:00

abstract：:Oligonucleotide primers used to amplify target DNA regions via PCR should meet certain design criteria to maximize the potential for efficient priming. The Random Oligonucleotide Construction Kit (ROCK), a spreadsheet-based program that runs under Microsoft Excel 97 or later version for Microsoft Windows, was develope...

journal_title：BioTechniques

authors： Strain SR,Chmielewski JG

更新日期：2001-06-01 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：:A major problem in analyzing forensic casework samples is the presence of genetic material from more than one individual in the material evidence. For instance, in sexual assault cases the evidence (vaginal swabs) usually contains a majority of vaginal epithelial cells and varying amounts of sperm cells from the perpe...

journal_title：BioTechniques

authors： Allen M,Saldeen T,Gyllensten U

更新日期：1995-09-01 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：:The possibility to miniaturize and parallelize biological assays has a great impact on the development of biomedical technologies. Here, we describe a simple, miniaturized, and parallelized method employing entire cells from different cell lines displaying a protein of interest on their surface, which were immobilized...

journal_title：BioTechniques

authors： Schwenk JM,Stoll D,Templin MF,Joos TO

更新日期：2002-12-01 00:00:00

abstract：:We report a simple and inexpensive method to quantitate loading and transfer of RNA and to examine RNA integrity for use with Northern hybridization. This technique involves quantitation by computer-assisted video densitometry of a negative photograph of 28S and 18S rRNA from ethidium bromide-stained RNA fixed to nylo...

journal_title：BioTechniques

authors： Correa-Rotter R,Mariash CN,Rosenberg ME

更新日期：1992-02-01 00:00:00

abstract：:Microarray technologies have made possible comprehensive analyses of nucleic acid sequence and expression. However, the technology to obtain efficiently high-quality RNA and DNA suitable for array analysis from purified populations of neoplastic cells from human tissues has not been well addressed. Microdissection can...

journal_title：BioTechniques

authors： Barrett MT,Glogovac J,Prevo LJ,Reid BJ,Porter P,Rabinovitch PS

更新日期：2002-04-01 00:00:00

abstract：:We describe a convenient PCR-based protocol for in vitro recombination of homologous genes, thereby minimizing the rate of associated point mutations. High-fidelity recombination conditions were obtained using Vent DNA polymerase, which, in contrast to Taq DNA polymerase, shows significant proofreading activity and ra...

journal_title：BioTechniques

authors： Ninkovic M,Dietrich R,Aral G,Schwienhorst A

更新日期：2001-03-01 00:00:00

abstract：:Quantitative real-time PCR has become a popular method to analyze and quantify changes in the copy number of mitochondrial DNA (mtDNA), and nuclear DNA (nDNA) is often used as an endogenous reference for mtDNA abundance. In our experience, using nDNA as a reference is problematic, due to differences in the extraction ...

journal_title：BioTechniques

authors： Myers MB,Mittelstaedt RA,Heflich RH

更新日期：2009-10-01 00:00:00

abstract：:Silencing of mammalian gene expression by RNA interference (RNAi) technology can be achieved using small interfering RNA (siRNA) or short hairpin RNA (shRNA). However, the relative effectiveness of these two approaches is not known. It is also not clear whether gene-specific shRNA transcribed from an RNA polymerase II...

journal_title：BioTechniques

authors： Ling X,Li F

更新日期：2004-03-01 00:00:00

abstract：:Three-dimensional collagen gel contraction is the standard assay utilized for functionally quantifying a variety of cell types, in particular smooth muscle cells (SMCs) and myofibroblasts. Here, we have developed a method to effectively reduce the three-dimensional parameters of the standard collagen gel into a single...

journal_title：BioTechniques

authors： Ilagan R,Guthrie K,Quinlan S,Rapoport HS,Jones S,Church A,Basu J,Ludlow J

更新日期：2010-02-01 00:00:00