Fast separations of plasmid DNA using discontinuous gradients in the preparative ultracentrifuge.

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：:Radiolabeling of native proteins conventionally has required iodination using 125Iodine (125I). Although radioiodination can result in high specific activity, there are several drawbacks in the use of 125I (e.g., radiological hazards and short half-life). 14C-Methylamine-glutaraldehyde conjugation to proteins offers a...

journal_title：BioTechniques

authors： Moore PL,Damelin LH,Harrison TJ

更新日期：2003-08-01 00:00:00

abstract：:Fluorescent live imaging of cells and embryos at subcellular resolution poses significant challenges for biologists due to morbidity and mortality ensuing from phototoxicity. Here we report the use of a spinning-disk confocal microscope to image mouse and bovine preimplantation embryos without impairing their developm...

journal_title：BioTechniques

authors： Ross PJ,Perez GI,Ko T,Yoo MS,Cibelli JB

更新日期：2006-12-01 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：:The use of AlphaScreen® detection has allowed researchers to examine a wide variety of molecular interactions for use in high-throughput screening. However, the cost of Alpha reagents can often be prohibitory for extended screening campaigns or for young investigators with limited funding. To reduce assay costs, many ...

journal_title：BioTechniques

authors： Veloria JR,Devkota AK,Cho EJ,Dalby KN

更新日期：2018-04-01 00:00:00

abstract：:We describe here a simple and efficient transfection method for transient expression of cloned genes in cell lines and primary cultured cells. The method involves the use of DEAE-dextran to target DNA to the cellular endocytotic pathway and the use of a human adenovirus to ensure efficient lysis of endosomal vesicles....

journal_title：BioTechniques

authors： Forsayeth JR,Garcia PD

更新日期：1994-08-01 00:00:00

abstract：:Gene expression profiles may offer more or additional information than classic morphologic- and histologic-based tumor classification systems. Because the number of tissue samples examined is usually much smaller than the number of genes examined, efficient data reduction and analysis methods are critical. In this rep...

journal_title：BioTechniques

authors： Xiong M,Jin L,Li W,Boerwinkle E

更新日期：2000-12-01 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：:We describe a simple and efficient system for epitope mapping by cloning random gene fragments into a specially designed gIIIp-based phage display vector. DNA encoding the antigen of interest is PCR-amplified and partially digested with DNaseI to generate 50-150-bp-long fragments, which are polished with T4 DNA polyme...

journal_title：BioTechniques

authors： Gupta S,Arora K,Sampath A,Khurana S,Singh SS,Gupta A,Chaudhary VK

更新日期：1999-08-01 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：:The analysis of DNA restriction fragment length polymorphisms by Southern blot hybridization requires that sufficient quantities of high molecular weight genomic DNA be extracted from biological specimens. Prior to analysis, it is necessary to determine the quantity and quality of the extracted DNA. For many applicati...

journal_title：BioTechniques

authors： Waye JS,Presley LA,Budowle B,Shutler GG,Fourney RM

更新日期：1989-09-01 00:00:00

abstract：:Sequencing of the human genome in combination with computational annotation has provided tremendous data on predicted genes. However, for most of them, no corresponding cDNAs are available yet. Furthermore, even where cDNA clones were obtained, gene transcripts often have many different splice variants that are not co...

journal_title：BioTechniques

authors： Mitani Y,Nakayama T,Harbers M,Hayashizaki Y

更新日期：2004-07-01 00:00:00

abstract：:As more and more researchers are examining proteins that are available only in extremely limited quantities, i.e., cellular extracts or genetic engineering products, it is critical to utilize staining methods that maximize sensitivity. The protocol we describe here--double staining of polyacrylamide electrophoresis ge...

journal_title：BioTechniques

authors： Ross M,Peters L

更新日期：1990-11-01 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：:beta-galactosidase (beta-gal), the product of the E. coli LacZ gene, has been used extensively as a reporter in numerous systems. Until recently, the most commonly used method of detecting beta-gal reporter enzymatic activity was a colormetric assay based on the cleavage of the beta-gal substrate 5-bromo-4-chloro-3-in...

journal_title：BioTechniques

authors： Nazarenko DA,Dertinger SD,Gasiewicz TA

更新日期：2001-04-01 00:00:00

abstract：:Several DNA extraction methods have been reported for use with digesta or fecal samples, but problems are often encountered in terms of relatively low DNA yields and/or recovering DNA free of inhibitory substances. Here we report a modified method to extract PCR-quality microbial community DNA from these types of samp...

journal_title：BioTechniques

authors： Yu Z,Morrison M

更新日期：2004-05-01 00:00:00

abstract：:Identification of nucleotides used for RNA chain initiation or for contacting DNA binding proteins is basic to our understanding of gene regulation. Normally, a radioactive primer is used to copy RNA or DNA. The polymerase extension stops at free ends of mRNA (as in promoter mapping) or at the position of template cle...

journal_title：BioTechniques

authors： Fekete RA,Miller MJ,Chattoraj DK

更新日期：2003-07-01 00:00:00

abstract：:In this study, we have developed an air-interface culture system in which guinea pig tracheobronchial epithelial (GPTE) cells rapidly form tight monolayers. Enzymatically isolated GPTE cells were plated on collagen-treated polycarbonate microporous cell culture inserts at a density of 10(6) cells/cm2 (day 0). Bioelect...

journal_title：BioTechniques

authors： Robison TW,Dorio RJ,Kim KJ

更新日期：1993-09-01 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：: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：:WE present a rapid procedure based on the polymerase chain reaction for generation of double-stranded DNA templates suitable for in vitro transcription by T3 or T7 RNA polymerase. DNA fragments cloned into a phage promoter vector are amplified together with a flanking promoter to provide functional templates. Extensio...

journal_title：BioTechniques

authors： Weier HU,Rosette C

更新日期：1990-03-01 00:00:00

abstract：:Procedures utilizing Chelex 100 chelating resin have been developed for extracting DNA from forensic-type samples for use with the PCR. The procedures are simple, rapid, involve no organic solvents and do not require multiple tube transfers for most types of samples. The extraction of DNA from semen and very small blo...

journal_title：BioTechniques

authors： Walsh PS,Metzger DA,Higuchi R

更新日期：1991-04-01 00:00:00

abstract：:A low order neural network-based filter was designed as a rapid screening agent for single-spanning transmembrane regions in an integrated informatics system. A rapid screening algorithm was seen as a compromise between costly structure-specific techniques and simple rules that gave a high false-positive rate for cDNA...

journal_title：BioTechniques

authors： Huang GM,Farkas J,Hood L

更新日期：1996-12-01 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

abstract：:Under-agarose chemotaxis has been used previously to assess the ability of neutrophils to respond to gradients of chemoattractant. We have adapted this assay to the chemotactic movement of Dictyostelium amoebae in response to folic acid. Troughs are used instead of wells to increase the area along which the cells can ...

journal_title：BioTechniques

authors： Laevsky G,Knecht DA

更新日期：2001-11-01 00:00:00

abstract：:Stable cell transfection is used for the expression of exogenous genes or cDNAs in eukaryotic cells. Selection of these transfectants requires a dominant selectable marker. A variety of such markers has been identified and is currently in use. However, many of these are not suitable for all cell types or require uniqu...

journal_title：BioTechniques

authors： Baccam M,Huberman E

更新日期：2003-06-01 00:00:00

abstract：:Exposing cells to brief, high-intensity electrical field pulses can lead to the permeabilization of their plasma membranes. This electro-induced permeated state of the cell membrane is reversible and leads to cell fusion; i.e., the electropermeabilized state if fusogenic. The size of cells intended for fusing, however...

journal_title：BioTechniques

authors： Rols MP,Dahhou F,Teissié J

更新日期：1994-10-01 00:00:00

abstract：:Genome sequencing projects are either based on whole genome shotgun (WGS) or on a BAC-by-BAC strategy. Although WGS is in most cases the preferred choice, sometimes the BAC-by-BAC approach may be better because it requires a much simpler assembly process. Furthermore, when the study is limited to specific regions of t...

journal_title：BioTechniques

authors： Todesco S,Campagna D,Levorin F,D'Angelo M,Schiavon R,Valle G,Vezzi A

更新日期：2008-01-01 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：: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