Simplified DGS procedure for large-scale genome structural study.

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：:We propose two methods for characterizing the spatio-temporal behavior of cell populations in culture. The first method, image auto-correlation microscopy (IACM), allows us to characterize the variation in the number of objects as a function of time, thus enabling the quantification of the clustering properties of cel...

journal_title：BioTechniques

authors： Bonnet N,Delavoie F,Zahm JM

更新日期：2007-07-01 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：:Many DNA binding proteins are known to regulate gene expression. When that binding is altered, a disease state can result. A common method for measuring DNA binding, namely electrophoretic mobility shift assay (EMSA) is often used but it is not amenable to rapid screening of many samples. As an alternative method, we ...

journal_title：BioTechniques

authors： Gal S,Cook JR,Howells L

更新日期：2006-09-01 00:00:00

abstract：:Specific amplification of nucleic acid sequences by PCR has been extensively used for the detection of gene rearrangements and gene expression. Although successful amplification of DNA sequences has been carried out with DNA prepared from formalin-fixed, paraffin-embedded (FFPE) tissues, there are only a few reports r...

journal_title：BioTechniques

authors： Finke J,Fritzen R,Ternes P,Lange W,Dölken G

更新日期：1993-03-01 00:00:00

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：:prfectBLAST is a multiplatform graphical user interface (GUI) for the stand-alone BLAST+ suite of applications. It allows researchers to do nucleotide or amino acid sequence similarity searches against public (or user-customized) databases that are locally stored. It does not require any dependencies or installation a...

journal_title：BioTechniques

authors： Santiago-Sotelo P,Ramirez-Prado JH

更新日期：2012-11-01 00:00:00

abstract：:Recombinant retroviral vectors usually encode the genes of interest in place of the viral structural genes, which must be provided in trans. These viruses are therefore defective for replication: infected cells cannot produce progeny virus. However, in some cases it may be desirable to generate virus from an infected ...

journal_title：BioTechniques

authors： Corbley MJ,Cherington V,Feig LA,Cooper GM,Roberts TM

更新日期：1994-12-01 00:00:00

abstract：:High stabilities of reporter proteins and their messenger RNAs (mRNAs) interfere with the detection of rapid transient changes in gene expression, such as transcriptional blocks posed by genotoxic DNA lesions. We have modified a green fluorescent protein (GFP) gene within the episomal pMARS vector by addition of a fra...

journal_title：BioTechniques

authors： Kitsera N,Khobta A,Epe B

更新日期：2007-08-01 00:00:00

abstract：:The two-hybrid system in Saccharomyces cerevisiae is a genetic approach for the detection of of protein-protein interactions in vivo. This technology relies on the the activity of separated DNA-binding and transactivation domains of specific transcription factors to reconstitute an active transcription factor complex ...

journal_title：BioTechniques

authors： Mayer G,Launhardt H,Munder T

更新日期：1999-07-01 00:00:00

abstract：:Scanning conductance microscopy investigations were carried out in air on human chromosomes fixed on pre-fabricated SiO2 surfaces with a backgate. The point of the investigation was to estimate the dielectric constant of fixed human chromosomes in order to use it for microfluidic device optimization. The phase shift c...

journal_title：BioTechniques

authors： Clausen CH,Lange JM,Jensen LB,Shah PJ,Dimaki MI,Svendsen WE

更新日期：2008-02-01 00:00:00

abstract：:We have compared RNA polymerase promoter activities in PCR-generated DNA fragments for use in the in vitro transcription of cRNA probes. Sense oligonucleotide primers, specific for the mouse acidic fibroblast growth factor gene, were synthesized with 5' extensions containing promoter sequences for the T7, T3 and SP6 R...

journal_title：BioTechniques

authors： Logel J,Dill D,Leonard S

更新日期：1992-10-01 00:00:00

abstract：:A consensus peptide sequence, QSYP, appears as an artifact during the mapping of monoclonal antibodies (MAbs) using a random peptide phage display library. Phage bearing this QSYP sequence were independently selected by four different laboratories screening separate MAb preparations with the same phage library. In eac...

journal_title：BioTechniques

authors： Jacobs JM,Bailey BW,Burritt JB,Morrison SG,Morrison RP,Dratz EA,Jesaitis AJ,Teintze M

更新日期：2003-01-01 00:00:00

abstract：:A critical issue in transfection or co-transfection experiments is to define the appropriate controls. In most cases, a corresponding empty vector is used as one control. We report a paradoxical effect of empty mammalian expression vectors on different reporters. We have found that different empty vectors can inhibit ...

journal_title：BioTechniques

authors： Hu Q,Suzuki K,Hirschler-Laszkiewicz I,Rothblum LI

更新日期：2002-07-01 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：:We describe a simple and inexpensive method of performing sequencing reactions for 24 single-strand M13 DNA clones in microtiter plates. To simplify elevated temperature incubations during sequencing reactions, two heating blocks were designed to accommodate microtiter plates and fit within common laboratory heating m...

journal_title：BioTechniques

authors： Koop BF,Wilson RK,Chen C,Halloran N,Sciammis R,Hood L,Lindelien JW

更新日期：1990-07-01 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：:For immunological research on the human cytomegalovirus (HCMV), a virus that combines the broad cell tropism of clinical isolates, efficient replication in cell culture, the complete set of MHC-I modulator genes, and suitability for genetic engineering is desired. Here, we aimed to generate a genetically complete deri...

journal_title：BioTechniques

authors： Sampaio KL,Weyell A,Subramanian N,Wu Z,Sinzger C

更新日期：2017-11-01 00:00:00

abstract：:The Drosophila heart has gained considerable traction as a model of cardiac development and physiology. Previously we described a semiautomatic optical heartbeat analysis (SOHA) method for quantifying functional parameters from the fly heart that facilitated its use as an organ system and disease model. Here we presen...

journal_title：BioTechniques

authors： Cammarato A,Ocorr S,Ocorr K

更新日期：2015-02-01 00:00:00

abstract：:The increase of information in biology makes it difficult for researchers in any field to keep current with the literature. The MEDLINE database of scientific abstracts can be quickly scanned using electronic mechanisms. Potentially interesting abstracts can be selected by matching words joined by Boolean operators. H...

journal_title：BioTechniques

authors： Perez-Iratxeta C,Keer HS,Bork P,Andrade MA

更新日期：2002-06-01 00:00:00

abstract：:Data are presented illustrating the optimum concentration range of reverse transcription PCR-generated products under 500 bp for accurate base calling with direct automated DNA sequencing. A 357-bp fragment of the human estrogen receptor, which includes the DNA binding domain of the protein, was used as a representati...

journal_title：BioTechniques

authors： Hyder SM,Hu C,Needleman DS,Sonoda Y,Wang XY,Baker VV

更新日期：1994-09-01 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：:It is difficult to isolate rare, PCR-quality DNA from specimens containing large quantities of nonspecific DNA from multiple sources (heterogeneous DNA). Extracting human DNA from stool for colorectal cancer (CRC) screening tests exemplifies this technically challenging sample preparation problem. The stool matrix is ...

journal_title：BioTechniques

authors： Kent Moore J,Smith JA,Whitney DH,Durkee KH,Shuber AP

更新日期：2008-03-01 00:00:00

abstract：:A method was developed for the detection of bacterial mRNAs using reverse transcriptase followed by the polymerase chain reaction (PCR) and Southern blot analysis. The method involves brief inhibition of protein synthesis with chloramphenicol, followed by reverse transcription, PCR amplification of cDNA and Southern b...

journal_title：BioTechniques

authors： Mahbubani MH,Bej AK,Miller RD,Atlas RM,DiCesare JL,Haff LA

更新日期：1991-01-01 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：:Recombinant alphaviruses have been used as vehicles for delivery and expression of heterologous genes in mammalian, avian and insect cell lines. We have used a Sindbis replicon virus (Sinreplac) able to express the E. coli lacZ gene to compare the efficiency of transduction in one insect, six mammalian cell lines and ...

journal_title：BioTechniques

authors： Corsini J,Traul DL,Wilcox CL,Gaines P,Carlson JO

更新日期：1996-09-01 00:00:00

abstract：:Complete brain fixation can be achieved with transthoracic cardiac infusion without thoracotomy. Light and electron microscopy tissue sections reveal preservation of cytoplasmic and nuclear structure at all magnification levels. Punched samples were obtained from the fixed tissue specimens in precisely localized areas...

journal_title：BioTechniques

authors： Eichenbaum KD,Eichenbaum JW,Fadiel A,Miller DC,Demir N,Naftolin F,Stern A,Pevsner PH

更新日期：2005-10-01 00:00:00

abstract：:The transcription factor GATA-1 is a zinc finger DNA-binding protein essential for the development of red blood cells. When we expressed different regions of the zinc finger domain in bacteria using an isopropyl-beta-D-thiogalactoside (IPTG) inducible system, growth of bacteria harboring the active DNA-binding domain ...

journal_title：BioTechniques

authors： Trudel P,Provost S,Massie B,Chartrand P,Wall L

更新日期：1996-04-01 00:00:00

abstract：:High-throughput approaches for gene cloning and expression require the development of new nonstandard tools for molecular biologists and biochemists. We introduce a Web-based tool to design primers specifically for the generation of expression clones for both laboratory-scale and high-throughput projects. The applicat...

journal_title：BioTechniques

authors： Yoon JR,Laible PD,Gu M,Scott HN,Collart FR

更新日期：2002-12-01 00:00:00

abstract：:Tissue engineering scaffolds have emerged as a powerful tool within regenerative medicine. These materials are being designed to create environments that promote regeneration through a combination of: (i) scaffold architecture, (ii) the use of scaffolds as vehicles for transplanting progenitor cells, and/or (iii) loca...

journal_title：BioTechniques

authors： Boehler RM,Graham JG,Shea LD

更新日期：2011-10-01 00:00:00