Simultaneous mutagenesis of multiple sites: application of the ligase chain reaction using PCR products instead of oligonucleotides.

abstract：:Random mutagenesis followed by an in vitro selection procedure was shown to be capable of identifying important bases of the hairpin ribozyme for cleavage of an RNA target sequence. The selection scheme enriched the RNA population for those molecules capable of efficient site-specific self-cleavage in the absence of l...

journal_title：BioTechniques

authors： Siwkowski A,Humphrey M,De-Young MB,Hampel A

更新日期：1998-02-01 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：: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：:We have altered the antibiotic resistance of the reporter plasmids and the pJG4-5 activation-domain and pEG202 DNA binding-domain plasmids used in the Brent interaction trap/two-hybrid system. These plasmids were each previously ampicillin-resistant, resulting in an inefficient purification of any one plasmid from a y...

journal_title：BioTechniques

authors： Watson MA,Buckholz R,Weiner MP

更新日期：1996-08-01 00:00:00

abstract：:A modification of the TRI Reagent procedure has been elaborated for isolation of RNA from polysaccharide- and proteoglycan-rich material. In the modified procedure, RNA is precipitated from the aqueous phase by the combined action of isopropanol and a high-salt concentration. Under these conditions, RNA is effectively...

journal_title：BioTechniques

authors： Chomczynski P,Mackey K

更新日期：1995-12-01 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：: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：: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：:Directed evolution is poised to change small molecule discovery and provide greater access to "drug space." Sarah Webb looks into the evolving drug discovery landscape. ...

journal_title：BioTechniques

authors： Webb S Ph D

更新日期：2017-12-01 00:00:00

abstract：:We present a tool for dispensing very low volumes (20 nL or more) of ultra high viscosity (UHV) medical-grade alginate hydrogels. It uses a modified piezo-driven micrometering valve, integrated into a versatile system that allows fast prototyping of encapsulation procedures and scaffold production. Valves show excelle...

journal_title：BioTechniques

authors： Gepp MM,Ehrhart F,Shirley SG,Howitz S,Zimmermann H

更新日期：2009-01-01 00:00:00

abstract：:Recombinant vaccinia viruses (rVACVs) are promising antigen-delivery systems for vaccine development that are also useful as research tools. Two common methods for selection during construction of rVACV clones are (i) co-insertion of drug resistance or reporter protein genes, which requires the use of additional selec...

journal_title：BioTechniques

authors： Liu L,Cooper T,Eldi P,Garcia-Valtanen P,Diener KR,Howley PM,Hayball JD

更新日期：2017-04-01 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：:An improved Huffman coding method for information storage in DNA is described. The method entails the utilization of modified unambiguous base assignment that enables efficient coding of characters. A plasmid-based library with efficient and reliable information retrieval and assembly with uniquely designed primers is...

journal_title：BioTechniques

authors： Ailenberg M,Rotstein O

更新日期：2009-09-01 00:00:00

abstract：:Human multiple tissue Western (MTW) blots are premade immunoblots prepared using proteins isolated from adult human tissue. The proteins are isolated from whole tissue homogenates under conditions designed to minimize proteolysis and to ensure maximal representation of tissue-specific proteins. Sodium dodecyl sulfate ...

journal_title：BioTechniques

authors： Kain SR,Mai K,Sinai P

更新日期：1994-11-01 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：:Much effort has been directed toward the isolation and characterization of homeobox cDNAs from numerous cell types because they encode transcription factors important to many cellular processes, including pattern formation in the embryo, cell growth and cell differentiation. Many novel homeobox cDNAs have been isolate...

journal_title：BioTechniques

authors： Gorski DH,LePage DF,Walsh K

更新日期：1994-05-01 00:00:00

abstract：:We have developed a novel method for the rapid preparation of large quantities of 3' end-labeled single-stranded (ss) DNA (ssDNA) probes for transcript mapping. A recombinant phagemid vector containing the probe sequence was used to raise large quantities of ssDNA. Based on the DNA sequence of the probe, an oligonucle...

journal_title：BioTechniques

authors： Silk GW,Wu M

更新日期：1993-05-01 00:00:00

abstract：:A general method for solid-phase gene assembly on streptavidin-coated magnetic beads has been developed. The introduction of biotin in the 5'-end of the initiation oligonucleotide enables anchoring to the bead by means of the streptavidin-biotin interaction. The immobilization of one oligonucleotide enables controlled...

journal_title：BioTechniques

authors： Ståhl S,Hansson M,Ahlborg N,Nguyen TN,Liljeqvist S,Lundeberg J,Uhlén M

更新日期：1993-03-01 00:00:00

abstract：:Automated DNA sequencing requires the intensive use of computers to handle the large amount of data taken. When a computer failure occurs and the data are no longer accessible, all the expense and effort that went into the sequencing experiment is lost. By using the data storage architecture of Macintosh computers to ...

journal_title：BioTechniques

authors： O'Brien KM,Fondon JW 3rd,Evans GA,Garner HR

更新日期：1997-06-01 00:00:00

abstract：:Advances in in vitro display and protein engineering yield therapeutics with affinities in the picomolar range. The Gyrolab® microfluidics platform uses the kinetic exclusion assay principle to measure subnanomolar solution affinities. This work describes application of the Gyrolab solution affinity module and the new...

journal_title：BioTechniques

authors： Dai Z,Juneja J,Schneeweis L,Cohen D,Marsilio F,Morin P,DasGupta R

更新日期：2020-09-01 00:00:00

abstract：:A pipeline has been created for the characterization of Arabidopsis thaliana mutants by generating flanking sequence tags (FSTs) and optimized for economic, high-throughput production. The GABI-Kat collection of T-DNA mutagenized A. thaliana plants was used as a source of independent transgenic lines. The pipeline inc...

journal_title：BioTechniques

authors： Strizhov N,Li Y,Rosso MG,Viehoever P,Dekker KA,Weisshaar B

更新日期：2003-12-01 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：:In order to study quantitative gene expression with Northern blots, it is important to have an internal standard that can be used to verify even loading or to correct for uneven loading between lanes. In this study it is shown that two-dimensional quantitation of ethidium bromide-intercalated 28S rRNA fluorescence can...

journal_title：BioTechniques

authors： Bonini JA,Hofmann C

更新日期：1991-12-01 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：:Sequence-based methods for transcriptome characterization have typically relied on generation of either serial analysis of gene expression tags or expressed sequence tags. Although such approaches have the potential to enumerate transcripts by counting sequence tags derived from them, they typically do not robustly su...

journal_title：BioTechniques

authors： Morin R,Bainbridge M,Fejes A,Hirst M,Krzywinski M,Pugh T,McDonald H,Varhol R,Jones S,Marra M

更新日期：2008-07-01 00:00:00

abstract：:An assay is described in which an oligonucleotide probe is specifically digested by lambda exonuclease only when it is annealed to its complementary sequence. In this assay, a cycling effect occurs whereby a small amount of target sequence acts as a specific co-factor in the enzymatic degradation of a larger number of...

journal_title：BioTechniques

authors： Copley CG,Boot C

更新日期：1992-12-01 00:00:00

abstract：:The yeast two-hybrid system has been used to characterize many protein-protein interactions. A two-hybrid system for E. coli was constructed in which one hybrid protein bound to a specific DNA site recruits another to an adjacent DNA binding site. The first hybrid comprises a test protein, the bait, fused to a chimeri...

journal_title：BioTechniques

authors： Hays LB,Chen YS,Hu JC

更新日期：2000-08-01 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：:Real-time or quantitative loop-mediated isothermal amplification (qLAMP) is a promising technique for the accurate detection of pathogens in organisms and the environment. Here we present a comparative study of the performance of six fluorescent intercalating dyes-SYTO-9, SYTO-13, SYTO-82, SYBR Green I, SYBR Gold, Eva...

journal_title：BioTechniques

authors： Oscorbin IP,Belousova EA,Zakabunin AI,Boyarskikh UA,Filipenko ML

更新日期：2016-07-01 00:00:00

abstract：:The conventional method for cloning a DNA fragment is to insert it into a vector and ligate it. Although this method is commonly used, it is labor intensive because the ratio and concentrations of the DNA insert and the vector need optimizing. Even then, the resultant library is often plagued with unwanted plasmids th...

journal_title：BioTechniques

authors： Miyazaki K,Takenouchi M

更新日期：2002-11-01 00:00:00