Rapid diagnostic assays in the genomic biology era: detection and identification of infectious disease and biological weapon agents.

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：: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：: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：:Next-generation sequencing (NGS) of multigene panels performed for genetic clinical diagnostics requires 100% coverage of all targeted genes. In the genetic diagnostics laboratory, coverage gaps are typically filled with Sanger sequencing after NGS data are collected and analyzed. Libraries prepared using the hybridiz...

journal_title：BioTechniques

authors： Coonrod EM,Durtschi JD,VanSant Webb C,Voelkerding KV,Kumánovics A

更新日期：2014-10-01 00:00:00

abstract：:An easy, sensitive and direct fluorescent immunodetection method for proteins is described using the new fluorochrome PBXL-1 imaged with the FMBIO II Laser Scanning Imaging System. PBXL-1 is derived from a protein supra-molecular complex that contains a large number of chromophores. This complex, the phycobilisome, is...

journal_title：BioTechniques

authors： Morseman JP,Moss MW,Zoha SJ,Allnutt FC

更新日期：1999-03-01 00:00:00

abstract：:The use of reverse transcription (RT) PCR for relative quantitation of gene transcripts relies on the reproducibility of the individual RT, PCR and product measurement steps. Semi-competitive RT-PCR (RT-cPCR) uses an internal competitor template in the PCR step to improve quantitation. We have surveyed the reproducibi...

journal_title：BioTechniques

authors： Hall LL,Bicknell GR,Primrose L,Pringle JH,Shaw JA,Furness PN

更新日期：1998-04-01 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：: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：: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：:The reliability of the PCR technique used to type two human variable number tandem repeats, that is, 3' to apolipoprotein B gene and locus D17S30, was examined using DNA samples of mixed human and microbial origin. Mixtures of human and microbial DNA were amplified, choosing microbes found commonly in the vagina. Tota...

journal_title：BioTechniques

authors： Lienert K,Fowler JC

更新日期：1992-08-01 00:00:00

abstract：:We describe a rapid, simple and inexpensive method for the isolation of DNA from blood clots suited for use in PCR. Our method is based on the lysing and nuclease-inactivating properties of guanidine thiocyanate together with the nucleic acid-binding properties of silica particles. Isolated DNA can be used for in vitr...

journal_title：BioTechniques

authors： Zeillinger R,Schneeberger C,Speiser P,Kury F

更新日期：1993-02-01 00:00:00

abstract：:A fully automated nucleic acid analysis system is described, which offers positive sample identification, improved sensitivity and reduced user interaction compared to conventional techniques. The system relies on the sequence-specific capture of DNA onto solid-phase particles, confirming product identity without the ...

journal_title：BioTechniques

authors： Brown A,Akinsanya AA,Barker SJ,Brophy M,Dobb AK,Doyle SM,Hudson IR,Minter SJ,Wraith MJ,Oultram JD

更新日期：1999-07-01 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：:We report a simple and inexpensive method to quantitate loading and transfer of RNA and to examine RNA integrity for use with Northern hybridization. This technique involves quantitation by computer-assisted video densitometry of a negative photograph of 28S and 18S rRNA from ethidium bromide-stained RNA fixed to nylo...

journal_title：BioTechniques

authors： Correa-Rotter R,Mariash CN,Rosenberg ME

更新日期：1992-02-01 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 simplified method for producing recombinant baculovirus for expression of foreign genes is described. The method utilizes insect cells infected with the wild-type virus before transfection with the plasmid transfer vector, instead of the standard procedure utilizing cotransfection with a plasmid and viral DNA. Recom...

journal_title：BioTechniques

authors： Goswami BB,Glazer RI

更新日期：1991-05-01 00:00:00

abstract：:Immunization with naked DNA was used to elicit chicken egg yolk antibodies (IgY). Layer hens were inoculated with plasmid DNA encoding the enhanced green fluorescent protein, the fusion protein of Newcastle disease virus, and VP2 of African horse sickness virus. IgY was extracted from egg yolks by polyethylene glycol ...

journal_title：BioTechniques

authors： Romito M,Viljoen GJ,Du Plessis DH

更新日期：2001-09-01 00:00:00

abstract：:We describe a multipurpose eukaryotic expression vector that incorporates the following features: restriction sites for in-frame insertion of cDNAs of interest between sequences encoding the glutathione-S-transferase (GST) and an oligohistidine element, allowing expression of the corresponding fusion proteins; a phosp...

journal_title：BioTechniques

authors： Chatton B,Bahr A,Acker J,Kedinger C

更新日期：1995-01-01 00:00:00

abstract：:Genomic analysis of circulating, cell-free DNA (cfDNA) is being used extensively for molecular diagnostics. Many approaches rely on the construction of cfDNA genomic libraries, targeted retrieval of specific genomic regions and analysis by next-generation DNA sequencing. Several steps during sample preparation require...

journal_title：BioTechniques

authors： Raymond CK

更新日期：2019-10-01 00:00:00

abstract：:We describe a general method for making template DNA for sequencing of PCR products. The procedure may be particularly useful for PCR products where minimal sequence information is known or as an alternative to primer walking when sequencing long PCR products. A cassette containing the hybridization site for the M13 s...

journal_title：BioTechniques

authors： Berg ES,Olaisen B

更新日期：1994-11-01 00:00:00

abstract：:We observed the presence of contaminating NADH oxidation activity in maltose binding protein (MBP) fusion proteins expressed in Escherichia coli and purified using conventional amylose resin-based affinity chromatography. This contaminating NADH oxidation activity was detectable with at least four different enzymes fr...

journal_title：BioTechniques

authors： Guo F,Zhu G

更新日期：2012-04-01 00:00:00

abstract：:When PCR is carried out in a polyacrylamide gel, each target molecule forms a molecular colony that comprises many copies of the original template. By counting the number of colonies, one can directly determine the target titer, with 100% of the DNA molecules and approximately 15% of the RNA molecules being detected. ...

journal_title：BioTechniques

authors： Chetverina HV,Samatov TR,Ugarov VI,Chetverin AB

更新日期：2002-07-01 00:00:00

abstract：:SCORE, a program for computer-assisted scoring of Southern blots of clone DNA, retains the use of expert human judgment while taking over much of the drudgery of the scoring task. The primary functions of the program are to help make an aligned overlay of the fluorescence gel image and the autoradiogram blot image, to...

journal_title：BioTechniques

authors： Cannon TM,Koskela RJ,Burks C,Stallings RL,Ford AA,Hempfner PE,Brown HT,Fickett JW

更新日期：1991-06-01 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：:Loop-mediated isothermal amplification (LAMP) is a rapid and reliable sequence-specific isothermal nucleic acid amplification technique. To date, all reported real-time detection methods for LAMP have been restricted to single targets, limiting the utility of this technique. Here, we adapted standard LAMP primers to c...

journal_title：BioTechniques

authors： Tanner NA,Zhang Y,Evans TC Jr

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 magnetic bead-based system for DNA isolation utilizing monodisperse beads was tested with the aim of producing a general approach for PCR-ready DNA. This commercially available system was originally designed for isolating PCR-ready DNA from human whole blood. We tested diverse organisms belonging to the major groups...

journal_title：BioTechniques

authors： Rudi K,Kroken M,Dahlberg OJ,Deggerdal A,Jakobsen KS,Larsen F

更新日期：1997-03-01 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：:We report a modification of a liquid hybridization method that serves as a rapid, sensitive alternative to Southern hybridization for the analysis of polymerase chain reaction (PCR) products. An aliquot of the completed PCR is mixed with an internally nested, end-labeled oligonucleotide probe, and one cycle of PCR is ...

journal_title：BioTechniques

authors： Parker JD,Burmer GC

更新日期：1991-01-01 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