A rapid and reliable method to create tandem arrays of short DNA sequences.

abstract：:A high-throughput, fluorescence-based helicase assay using molecular beacons is described. The assay is tested using the NS3 helicase encoded by the hepatitis C virus (HCV) and is shown to accurately monitor helicase action on both DNA and RNA. In the assay, a ssDNA oligonucleotide molecular beacon, featuring a fluore...

journal_title：BioTechniques

authors： Belon CA,Frick DN

更新日期：2008-10-01 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：: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：:Chitin from crustacean shells has often been used to isolate and purify plant lectins that have an affinity for poly-N-acetylglucosamine (poly-GlcNAc). When we used washed chitin from crab shells as an affinity medium to isolate a lectin from Pinus strobus L. (eastern white pine) ovules, we found that a substance havi...

journal_title：BioTechniques

authors： Whitmore FA

更新日期：1992-02-01 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：:Francisella tularensis is one of the most deadly bacterial agents, yet most of the genetic determinants of pathogenesis are still unknown. We have developed an efficient targeted mutagenesis strategy in the model organism F. tularensis subsp. novicida by utilizing universal priming of optimized antibiotic resistance c...

journal_title：BioTechniques

authors： Liu J,Zogaj X,Barker JR,Klose KE

更新日期：2007-10-01 00:00:00

abstract：:Biopharmaceutical products are of great importance in the treatment or prevention of many diseases and represent a growing share of the global pharmaceutical market. The usual technology for protein synthesis (cell-based expression) faces certain obstacles, especially with 'difficult-to-express' proteins. Cell-free pr...

journal_title：BioTechniques

authors： Chiba CH,Knirsch MC,Azzoni AR,Moreira AR,Stephano MA

更新日期：2021-01-20 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：: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：:Reverse transcription PCR (RT-PCR) represents a sensitive and powerful tool for analyzing RNA. While it has tremendous potential for quantitative applications, a comprehensive knowledge of its technical aspects is required. Successful quantitative RT-PCR involves correction for experimental variations in individual RT...

journal_title：BioTechniques

authors： Freeman WM,Walker SJ,Vrana KE

更新日期：1999-01-01 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：: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：: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：:A new genetic marker was created in which sequences from enhanced green fluorescent protein were fused to those of puromycin N-acetyl transferase. The resulting fusion protein (EGFP-puro) conferred both green fluorescence and resistance to puromycin when expressed in mammalian cells. The utility of EGFP-puro as a sele...

journal_title：BioTechniques

authors： Abbate J,Lacayo JC,Prichard M,Pari G,McVoy MA

更新日期：2001-08-01 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：:Based on the requirement of a glutathione-deficient mutant strain of Saccharomyces cerevisiae to take up external glutathione for growth on synthetic media, a simple agar diffusion test for quantitative detection of total glutathione from various sources was established. Glutathione concentrations can be reliably dete...

journal_title：BioTechniques

authors： Schmidt M,Grey M,Brendel M

更新日期：1996-11-01 00:00:00

abstract：:To establish a nonradioactive method for demonstrating HPV DNA in routinely treated smears of the uterine cervix (alcohol fixation, staining according to Papanicolaou, preservation), in situ hybridizations were carried out in HeLa and SiHa cells grown on slides. After detailed investigations, the sensitivity and speci...

journal_title：BioTechniques

authors： Heiles HB,Genersch E,Kessler C,Neumann R,Eggers HJ

更新日期：1988-11-01 00:00:00

abstract：:This paper reports a novel method for the identification of nucleic acid target sequences when these targets have high sequence identity. Homologous genes are currently identified by sequencing. We hypothesize that by primer extension in the presence of selected nucleotides, genes with similar sequence can be identifi...

journal_title：BioTechniques

authors： Sandhu GS,Kline BC,Bolander ME,Sarkar G

更新日期：1993-06-01 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：: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：:Subtractive hybridization has been widely used for the identification of differentially expressed genes. Here we describe a simple, sensitive strategy of subtractive hybridization that involves binding the driver poly(A)+ RNA pool to paramagnetic Dynabeads Oligo (dT)25. After hybridization with target cDNA, the molecu...

journal_title：BioTechniques

authors： Mészáros M,Morton DB

更新日期：1996-03-01 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：:The green fluorescent protein (GFP) from the jellyfish Aequorea victoria is rapidly becoming an important reporter molecule for monitoring gene expression and protein localization in vivo, in situ and in real time. GFP emits bright green light (lambda max = 509 nm) when excited with UV or blue light (lambda max = 395 ...

journal_title：BioTechniques

authors： Kain SR,Adams M,Kondepudi A,Yang TT,Ward WW,Kitts P

更新日期：1995-10-01 00:00:00

abstract：:Knob heterochromatin served as the model for the development of fluorescent chromosome in situ hybridization on maize meiotic chromosomes. The meiotic chromosomes were hybridized with a digoxigenin-labeled RNA probe of the knob repeat sequence that is a component of the morphologically determined knob heterochromatin....

journal_title：BioTechniques

authors： Makowski ER,Ruzin SE

更新日期：1994-02-01 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 method allowing the evaluation of the structure and the calculation of the volume of a biofilm, using an optical microscope, is proposed based on the linear relation between the intensity of a pixel in biofilm images grabbed on the x-y plane and the corresponding number of cells in the z direction, which allows the ...

journal_title：BioTechniques

authors： de Carvalho CC,da Fonseca MM

更新日期：2007-05-01 00:00:00

abstract：:We present a simple method for sequential chemiluminescent detections of two different DNA loci on a single Southern blot. First, an enzyme-linked DNA probe for a unique sequence is detected with a horse-radish peroxidase (HRP) substrate followed by the detection of another enzyme-linked DNA probe for a different uniq...

journal_title：BioTechniques

authors： Reddy LV,DeSilva R,Handley RS,Schaap AP,Akhavan-Tafti H

更新日期：1999-04-01 00:00:00

abstract：:Real-time PCR is becoming a preferred method for quantification of minute amounts of nucleic acids. To achieve the full potential of this technique, accurate and convenient models for post-PCR data analysis are required. In this study, three different models were chosen to quantify the definitive copy numbers of Cucum...

journal_title：BioTechniques

authors： Feng J,Zeng R,Chen J

更新日期：2008-06-01 00:00:00

abstract：:A head-to-tail trimer of the SV40 Bcl I-Bam H1 DNA fragment, specifying polyadenylation of RNA transcripts, was cloned as a cassette flanked by multiple restriction sites. Insertion of the trimer into several expression vectors efficiently prevented spurious expression of reporter genes resulting from transcriptional ...

journal_title：BioTechniques

authors： Maxwell IH,Harrison GS,Wood WM,Maxwell F

更新日期：1989-03-01 00:00:00

abstract：:Cycling probe technology (CPT) is a simple signal amplification method for the detection of specific target DNA sequences. CPT uses a chimeric DNA-RNA-DNA probe that is cut by RNase H when bound to its complementary target sequence. In this study, a hybridization assay was developed to detect biotinylated CPT products...

journal_title：BioTechniques

authors： Warnon S,Zammatteo N,Alexandre I,Hans C,Remacle J

更新日期：2000-06-01 00:00:00