New cloning vectors to facilitate quick allelic exchange in gram-negative bacteria.

abstract：:Naturstoff reagent A (diphenylboric acid 2-aminoethyl ester [DPBA]) has been used historically in plant science to observe polyphenolic pigments, such as flavonoids, whose fluorescence requires enhancement to be visible by microscopy. Flavonoids are common dietary constituents and are the focus of considerable attenti...

journal_title：BioTechniques

authors： Ferrara BT,Thompson EP

更新日期：2019-02-01 00:00:00

abstract：:A new configuration of the solid-support invasive cleavage reaction provides a small reaction-volume format for high-sensitivity discrimination of nucleic acid targets with single nucleotide differences. With target concentrations as low as 2 amol/assay, the solid-support invasive cleavage reaction clearly distinguish...

journal_title：BioTechniques

authors： Stevens PW,Rao KV,Hall JG,Lyamichev V,Neri BP,Kelso DM

更新日期：2003-01-01 00:00:00

abstract：:Reporter assays that use luciferase are widely employed for monitoring cellular events associated with gene expression. In general, firefly luciferase and Renilla luciferase are used for monitoring single gene expression. However, the expression of more than one gene cannot be monitored simultaneously by this system b...

journal_title：BioTechniques

authors： Nakajima Y,Kimura T,Sugata K,Enomoto T,Asakawa A,Kubota H,Ikeda M,Ohmiya Y

更新日期：2005-06-01 00:00:00

abstract：:A technique, Replacement PCR Mutagenesis, was developed to replace one immunoglobulin variable region (V) in a M13 phage cassette with a different, homologous V. This allows the use of the same mutagenesis and subsequent expression vectors for many V regions or V segments. The method combines PCR of V fragments and in...

journal_title：BioTechniques

authors： Near RI

更新日期：1992-01-01 00:00:00

abstract：:A variety of technical errors have arisen in data analysis when using cDNA or oligonucleotide microarrays. One of the most insidious problems is the saturation of the hybridization signal of high-abundant transcripts. This problem arises from the truncation of the laser fluorescence signal. When the hybridization sign...

journal_title：BioTechniques

authors： Hsiao LL,Jensen RV,Yoshida T,Clark KE,Blumenstock JE,Gullans SR

更新日期：2002-02-01 00:00:00

abstract：:Transfection of suspension cells has proven to be very difficult using conventional methods. Here, we present a simple and time-saving new transfection protocol wherein cell culture plates coated with chicken egg white are seeded with suspension cells prior to transfection. Our results demonstrate that coupling egg wh...

journal_title：BioTechniques

authors： Basiouni S,Fuhrmann H,Schumann J

更新日期：2012-08-01 00:00:00

abstract：:Pairs of primers flanking known miniTn10 transposon insertion sites were used to confirm the presence of the transposon in DNA isolated from Legionella pneumophila mutants. It was expected that the polymerase chain reaction products derived from the mutant template would be larger than those from the wild-type (WT) te...

journal_title：BioTechniques

authors： Viswanathan VK,Krcmarik K,Cianciotto NP

更新日期：1999-09-01 00:00:00

abstract：:A direct method to study essential genes is to construct conditional knock-down mutants by replacement of their native promoter by an inducible one. In Mycobacteria, replacement of an essential gene promoter with an anhydrotetracycline inducible one was successfully used but required a multi-step approach. In this wor...

journal_title：BioTechniques

authors： Texier P,Coddeville M,Bordes P,Genevaux P

更新日期：2018-09-01 00:00:00

abstract：:Cell-based immunizations are often used when membrane antigens are difficult to purify. To confirm that an antibody binding to the surface of a cell line is, in fact, binding to the desired antigen, FACS can be performed independently on two cell lines, a transfected cell line expressing the antigen of interest and a ...

journal_title：BioTechniques

authors： Yang XP,Gallo M,Ngan I,Nocerini M,Chen MM

更新日期：2002-03-01 00:00:00

abstract：:The ZEBRA protein is a transcriptional activator that induces expression of viral lytic genes in cells harboring latent Epstein-Barr virus (EBV). In this report it is shown that a derivative of ZEBRA that cannot activate transcription (Zd) can be used to detect and characterize activation domains. Three expression vec...

journal_title：BioTechniques

authors： Asković S,Baumann R

更新日期：1997-05-01 00:00:00

abstract：:In this special section of BioTechniques, we examine the role of rapid molecular technologies in the detection and identification of agents of infectious disease (ID) and biological weapons (BWs). Besides the threat posed by the global proliferation of BW technologies, there are numerous emerging and reemerging ID age...

journal_title：BioTechniques

authors： Peruski LF Jr,Peruski AH

更新日期：2003-10-01 00:00:00

abstract：:New technologies are needed that can diagnose cancer more rapidly and accurately. These technologies must also have the ability to identify the particular cellular abnormalities contributing to the malignancy, thus directing the appropriate treatments. Such technologies should permit absolute quantitation of specific ...

journal_title：BioTechniques

authors： Warren EN,Jiang J,Parker CE,Borchers CH

更新日期：2005-06-01 00:00:00

abstract：:Replication studies on human immunodeficiency virus 1 (HIV-1) rely on a few laboratory strains that are divergent from dominant HIV-1 subtypes in the epidemic. Several phenotypic differences between diverse HIV-1 isolates and subtypes could affect vaccine development and treatment, but this research field lacks robust...

journal_title：BioTechniques

authors： Dudley DM,Gao Y,Nelson KN,Henry KR,Nankya I,Gibson RM,Arts EJ

更新日期：2009-05-01 00:00:00

abstract：:The relative spatial distribution of cells in a solid tumor contributes to development of malignancy, yet the details of this process remain poorly understood. To elucidate these mechanisms, the ability to extract and analyze the entire DNA content of individual cells whose precise location in the tumor is known is re...

journal_title：BioTechniques

authors： Guo Y,Yang Y,Zhou J,Czajkowsky DM,Liu B,Shao Z

更新日期：2012-04-01 00:00:00

abstract：:To avoid the time-consuming reprobing process in hybridization analysis, signal-distinguishable probes (32P, 35S or antigenic hapten-labeled DNA) can be added to the same hybridization mixture. After hybridization, an unambiguous result can be obtained by differential or sequential autoradiography. ...

journal_title：BioTechniques

authors： Au LC,Chang KJ,Shih CM,Teh GW

更新日期：1994-04-01 00:00:00

abstract：:DNAdraw is a Macintosh program designed to prepare DNA and protein data for publication. In additional to providing standard ways of highlighting data, e.g., fonts, styles and shading, DNAdraw has special features for formatting sequence data and for handling aligned sequence data. Output for the program is to a PostS...

journal_title：BioTechniques

authors： Shapiro M

更新日期：1995-06-01 00:00:00

abstract：:Intrinsically disordered proteins (IDPs) are subject to post-translational modifications. This allows the same polypeptide to be involved in different interaction networks with different consequences, ranging from regulatory signalling networks to the formation of membrane-less organelles. We report a robust method fo...

journal_title：BioTechniques

authors： Brázda P,Šedo O,Kubíček K,Štefl R

更新日期：2019-07-01 00:00:00

abstract：:Duracryl is a mechanically strong and elastic acrylamide-based matrix, useful for a wide variety of electrophoretic applications. The matrix is stable as a refrigerated solution for one year. Upon addition of appropriate catalysts, Duracryl forms a polymer-reinforced polyacrylamide gel matrix suitable for electrophore...

journal_title：BioTechniques

authors： Patton WF,Lopez MF,Barry P,Skea WM

更新日期：1992-04-01 00:00:00

abstract：:PhDs, postdocs, fellows and newly appointed lecturers all fall into the category of early-career researchers (ECRs). The life of researchers in early stages of their career is far from straightforward. Funding is a huge barrier for many ECRs looking to make the next step in their career, but it's not the only challeng...

journal_title：BioTechniques

authors：

更新日期：2019-01-01 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：:In situ hybridization techniques typically employ chromogenic staining by enzymatic amplification to detect domains of gene expression. We demonstrate the previously unreported near infrared (NIR) fluorescence of the dark purple stain formed from the commonly used chromogens, nitro blue tetrazolium (NBT) and 5-bromo-4...

journal_title：BioTechniques

authors： Trinh le A,McCutchen MD,Bonner-Fraser M,Fraser SE,Bumm LA,McCauley DW

更新日期：2007-06-01 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：:The study of gene regulation in cardiac myocytes requires a reliable in vitro model. However, monolayer cultures used for this purpose are typically not exposed to electrical stimulation, though this has been shown to strongly affect cardiomyocyte gene expression. Based on pacemakers for clinical use, we developed an ...

journal_title：BioTechniques

authors： Martherus RS,Zeijlemaker VA,Ayoubi TA

更新日期：2010-01-01 00:00:00

abstract：:Radiolabeled proteins are used in a variety of laboratory applications as well as in radioimmunotherapy. This review focuses on methods that utilize genetic engineering to introduce exogenous phosphorylation sites into proteins. Protein kinase substrate sites can be introduced into target proteins to serve as tags for...

journal_title：BioTechniques

authors： Clark WA,Izotova L,Philipova D,Wu W,Lin L,Pestka S

更新日期：2002-10-01 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

abstract：:We describe a method for rapid radioactive labeling of PCR product. The method, employing the Klenow fragment of DNA polymerase I, consumes little product, requires no product purification and takes under 30 minutes. ...

journal_title：BioTechniques

authors： McDaniel TK,Huang Y,Yin J,Needleman SW,Meltzer SJ

更新日期：1991-08-01 00:00:00

abstract：:Methods have been developed to rapidly visualize the size distribution of third complementarity-determining regions (CDR3) in immunoglobulin (Ig) and T-cell receptor (TCR) molecules. DNA fragments spanning the Ig or TCR CDR3 are generated by PCR using primers at fixed positions in the variable and constant segments. T...

journal_title：BioTechniques

authors： Raaphorst FM,Tami J,Sanz IE

更新日期：1996-01-01 00:00:00

abstract：:Here we describe a convenient method to generate homologous recombinant baculoviral genomes in E. coli. The recombination takes place with the aid of recombination enzymes provided by the phage lambda Red system between a bacmid (a baculoviral genome that can replicate in bacteria) and a linear fragment. Proof of conc...

journal_title：BioTechniques

authors： Hou S,Chen X,Wang H,Tao M,Hu Z

更新日期：2002-04-01 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 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