High-efficiency transfection of suspension cell lines.

abstract：:There are little independent data available about how well single nucleotide polymorphism (SNP) genotyping technologies perform in the typical molecular genetics laboratory. We evaluated the utility and accuracy of a widely used technology, template-directed dye-terminator incorporation with fluorescence-polarization ...

journal_title：BioTechniques

authors： Akula N,Chen YS,Hennessy K,Schulze TG,Singh G,McMahon FJ

更新日期：2002-05-01 00:00:00

abstract：:Plasmid DNA can be purified in 20 min using a three-step CsCl gradient in a vertical tube rotor. Ultracentrifugation with discontinuous gradients yields DNA with purity comparable to that obtained with longer isopycnic separations and with columns. ...

journal_title：BioTechniques

authors： Dorin M,Bornecque CA

更新日期：1995-01-01 00:00:00

abstract：:The limiting detection signal for identification of human genetic markers, such as HLA-D and VNTR genes, was determined using DNA isolated from a series of decreasing numbers of lymphocytes carrying the target marker in the polymerase chain reaction (PCR). The PCR procedure was assembled by incorporating 32P-labeled d...

journal_title：BioTechniques

authors： McDaniel DO,Naftilan J,Barber WH

更新日期：1993-07-01 00:00:00

abstract：:The colony formation assay (CFA) is the gold standard for measuring the effects of cytotoxic agents on cancer cells in vitro; however, in its traditional 6-well format, it is a time-consuming assay, particularly when evaluating combination therapies. In the interest of increased efficiency, the 6-well CFA was converte...

journal_title：BioTechniques

authors： Katz D,Ito E,Lau KS,Mocanu JD,Bastianutto C,Schimmer AD,Liu FF

更新日期：2008-02-01 00:00:00

abstract：:Loop-mediated isothermal amplification (LAMP), a novel gene amplification method, enables the synthesis of larger amounts of both DNA and a visible byproduct--namely, magnesium pyrophosphate--without thermal cycling. A positive reaction is indicated by the turbidity of the reaction solution or the color change after a...

journal_title：BioTechniques

authors： Goto M,Honda E,Ogura A,Nomoto A,Hanaki K

更新日期：2009-03-01 00:00:00

abstract：:Transfection experiments involving mixed cell cultures pose special problems because of differential uptake and expression of DNA by different cell types. In order to evaluate the expression of transfected DNA in primary cultures of embryonic myocardial cells, we have used immunofluorescence microscopy to analyze expr...

journal_title：BioTechniques

authors： Antin PB,Mar JH,Ordahl CP

更新日期：1988-07-01 00:00:00

abstract：:Changes in cardiac structure that depart from normal have generally been termed "remodeling". Assessment of ventricular remodeling at the cellular level should include measurement of myocyte dimensions. A well-established and reliable method to assess myocyte remodeling uses isolated cells and the Coulter Counter/Chan...

journal_title：BioTechniques

authors： Said S,Tamura T,Gerdes AM

更新日期：1998-09-01 00:00:00

abstract：:A nonradioactive functional assay was developed to quantitate DNA binding proteins. The assay was designed to allow the use of 96-well microplates for high sample throughput. We show that the assay can measure sequence-specific DNA binding of purified proteins as well as DNA binding activity present in whole cell extr...

journal_title：BioTechniques

authors： Gubler ML,Abarzúa P

更新日期：1995-06-01 00:00:00

abstract：:Single-cell genomics (SCG) is a recently developed tool to study the genomes of unculturable bacterial species. SCG relies on multiple-strand displacement amplification (MDA), PCR, and next-generation sequencing (NGS); however, obtaining sufficient amounts of high-quality DNA from samples is a major challenge when per...

journal_title：BioTechniques

authors： He J,Du S,Tan X,Arefin A,Han CS

更新日期：2016-03-01 00:00:00

abstract：:Direct contact-based coculture of human dermal fibroblasts and epidermal keratinocytes has been a long-standing and challenging issue owing to different serum and growth factor requirements of the two cell types. Existing protocols employ high serum concentrations (up to 10% fetal bovine serum), complex feeder systems...

journal_title：BioTechniques

authors： Kadam S,Vandana M,Kaushik KS

更新日期：2020-11-01 00:00:00

abstract：:Treatment with 2 mM CuSO4 was used to induce a Drosophila melanogaster metallothionein (Mtn) promoter that had been cloned into a recombinant baculovirus. Careful study revealed that the Mtn promoter functioned as an inducible, if somewhat "leaky" promoter within the context of baculovirus-infected cells. In the proce...

journal_title：BioTechniques

authors： Lanier LM,Storm K,Shafaie A,Volkman LE

更新日期：1997-10-01 00:00:00

abstract：:A method for cloning single-stranded oligonucleotides in a plasmid vector has been developed. The method relies on ligation of the oligonucleotide into suitable restriction enzyme sites of the cloning vector such that the site at the 5' end has a 5' overhang [for example, a Bgl II site (A decreases GATCT)], and the si...

journal_title：BioTechniques

authors： Mounts P,Wu TC,Peden K

更新日期：1989-04-01 00:00:00

abstract：:To gain insightful information about the mechanisms through which genes are activated and repressed requires gene reporter systems that are sensitive, robust, and cost-effective. Although numerous reporter gene technologies are commercially available, none are as sophisticated and user-friendly as beta-lactamase (BLA)...

journal_title：BioTechniques

authors： Qureshi SA

更新日期：2007-01-01 00:00:00

abstract：:Genotyping fish larvae is a valuable technique for numerous fields of study. While methods for collecting DNA from early stage larvae have been published, a non-lethal, non-invasive genotyping protocol for hatchlings that is amenable to high-throughput approaches is desirable. Here, we describe a method to individuall...

journal_title：BioTechniques

authors： Espinoza GJ,Poland JM,Bremer JRA

更新日期：2017-10-01 00:00:00

abstract：:Integral membrane G protein-coupled receptors (GPCRs) compose the single most prolific class of drug targets, yet significant functional and structural questions remain unanswered for this superfamily. A primary reason for this gap in understanding arises from the difficulty of forming soluble, monodisperse receptor m...

journal_title：BioTechniques

authors： Leitz AJ,Bayburt TH,Barnakov AN,Springer BA,Sligar SG

更新日期：2006-05-01 00:00:00

abstract：:As an improved strategy for producing functional macromolecules by in vitro evolutionary optimization, we propose an automated machine that can process up to 960 samples in parallel. It consists of a 960-well PCR machine with special sealed plastic reaction vessels and appropriate handling devices. We show that the he...

journal_title：BioTechniques

authors： Schober A,Walter NG,Tangen U,Strunk G,Ederhof T,Dapprich J,Eigen M

更新日期：1995-04-01 00:00:00

abstract：:Sample automation and management is increasingly important as the number and size of population-scale and high-throughput projects grow. This is particularly the case in large-scale population studies where sample size is far outpacing the commonly used 96-well plate format. To facilitate management and transfer of sa...

journal_title：BioTechniques

authors： Cario CL,Witte JS

更新日期：2018-12-01 00:00:00

abstract：:Xer-cise is a technique using antibiotic resistance cassettes flanked by dif sites allowing spontaneous and accurate excision from bacterial chromosomes with a high frequency through the action of the cellular recombinase XerCD. Here, we report a significant improvement of Xer-cise in Mycobacteria. Zeocin resistance c...

journal_title：BioTechniques

authors： Boudehen YM,Wallat M,Rousseau P,Neyrolles O,Gutierrez C

更新日期：2020-02-01 00:00:00

abstract：:Single-stranded long oligonucleotide-based (50- to 70-mer) microarrays offer several advantages over conventional cDNA microarrays. These include the easy preparation of the probes, low cost of array production, and low cross-contamination during probe handling. However, the application of oligonucleotide microarrays ...

journal_title：BioTechniques

authors： Kaposi-Novak P,Lee JS,Mikaelyan A,Patel V,Thorgeirsson SS

更新日期：2004-10-01 00:00:00

abstract：:WE present a rapid procedure based on the polymerase chain reaction for generation of double-stranded DNA templates suitable for in vitro transcription by T3 or T7 RNA polymerase. DNA fragments cloned into a phage promoter vector are amplified together with a flanking promoter to provide functional templates. Extensio...

journal_title：BioTechniques

authors： Weier HU,Rosette C

更新日期：1990-03-01 00:00:00

abstract：:Recent computational and bioinformatics advances have enabled the efficient creation of novel biocatalysts by reducing amino acid variability at hot spot regions. To further expand the utility of this strategy, we present here a tool called Multi-site Degenerate Codon Analyzer (MDC-Analyzer) for the automated design o...

journal_title：BioTechniques

authors： Tang L,Wang X,Ru B,Sun H,Huang J,Gao H

更新日期：2014-06-01 00:00:00

abstract：:A new qualitative PCR product detection assay called competitive amplified single mutation detection by selective probe hybridization immunoassay (CASSI) was developed for genotyping the most common apolipoprotein E (apoE) polymorphisms. Single target DNA strands immobilized using biotin on streptavidin-coated micropl...

journal_title：BioTechniques

authors： Köhler T,Rost AK,Purschwitz K,Vondran S,Remke H,Wagner O,Richter V

更新日期：1998-07-01 00:00:00

abstract：:Thin spun-coat films (~4 nm thick) of graphene oxide (GO) constitute a versatile surface chemistry compatible with a broad range of technologically important sensor materials. Countless publications are dedicated to the nuances of surface chemistries that have been developed for sensors, with almost every material hav...

journal_title：BioTechniques

authors： Mulvaney SP,Stine R,Long NC,Tamanaha CR,Sheehan PE

更新日期：2014-07-01 00:00:00

abstract：:GC-rich DNA regions were PCR-amplified with Taq DNA polymerase using either the canonical set of deoxynucleoside triphosphates or mixtures in which the dCTP had been partially or completely replaced by its N4-methylated analog, N4-methyl-2'-deoxycytidine 5'-triphosphate (N4me-dCTP). In the case of a particularly GC-ri...

journal_title：BioTechniques

authors： Flores-Juárez CR,González-Jasso E,Antaramian A,Pless RC

更新日期：2016-10-01 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：:Identification of nucleotides used for RNA chain initiation or for contacting DNA binding proteins is basic to our understanding of gene regulation. Normally, a radioactive primer is used to copy RNA or DNA. The polymerase extension stops at free ends of mRNA (as in promoter mapping) or at the position of template cle...

journal_title：BioTechniques

authors： Fekete RA,Miller MJ,Chattoraj DK

更新日期：2003-07-01 00:00:00

abstract：:Comprehensive analysis of DNA methylation patterns is critical for understanding the molecular basis of many human diseases. While hundreds of PCR-based DNA methylation studies are published every year, the selection and implementation of appropriate methods for these studies can be challenging for molecular genetics ...

journal_title：BioTechniques

authors： Hernández HG,Tse MY,Pang SC,Arboleda H,Forero DA

更新日期：2013-10-01 00:00:00

abstract：:Exposing cells to brief, high-intensity electrical field pulses can lead to the permeabilization of their plasma membranes. This electro-induced permeated state of the cell membrane is reversible and leads to cell fusion; i.e., the electropermeabilized state if fusogenic. The size of cells intended for fusing, however...

journal_title：BioTechniques

authors： Rols MP,Dahhou F,Teissié J

更新日期：1994-10-01 00:00:00

abstract：:Several methods and computer programs have been developed for estimating the size of DNA fragments from gel electrophoresis. However, methods are lacking that may facilitate in optimization of gel conditions. In this article, a computer program called DNAOPT is described, which was developed to assist researchers in t...

journal_title：BioTechniques

authors： Raghava GP

更新日期：1995-02-01 00:00:00

abstract：:Gene expression analysis using high-density cDNA or oligonucleotide arrays is a rapidly emerging tool for transcriptomics, the analysis of the transcriptional state of a cell or organ. One of the limitations of current methodologies is the requirement of a relatively large amount of total or polyadenylated RNA as star...

journal_title：BioTechniques

authors： Scherer A,Krause A,Walker JR,Sutton SE,Serón D,Raulf F,Cooke MP

更新日期：2003-03-01 00:00:00