Laser-based microscopic approaches: application to cell signaling in environmental lung disease.

abstract：:The well-characterized cell line Chinese hamster ovary (CHO) has been used to produce numerous biopharmaceuticals and is an important tool for basic research. However, introducing foreign DNA into specially modified CHO cells such as DG44 and Lec 3.2.8.1 can sometimes be an arduous process. Here we show that the Flp-i...

journal_title：BioTechniques

authors： Soler DC,Young AE,Vahedi-Faridi A,McCormick TS

更新日期：2018-07-01 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：: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：:A procedure for amplification by PCR of reproducible allele markers for amplified fragment length polymorphism (Amp-FLP) analysis is presented. We have prepared markers for the allelic products of the VNTR loci D1S80 (MCT118) and D17S30 (YNZ22) and for the hypervariable VNTR locus close to the 3' end of the apolipopro...

journal_title：BioTechniques

authors： Sajantila A,Puomilahti S,Johnsson V,Ehnholm C

更新日期：1992-01-01 00:00:00

abstract：:High-throughput approaches for gene cloning and expression require the development of new nonstandard tools for molecular biologists and biochemists. We introduce a Web-based tool to design primers specifically for the generation of expression clones for both laboratory-scale and high-throughput projects. The applicat...

journal_title：BioTechniques

authors： Yoon JR,Laible PD,Gu M,Scott HN,Collart FR

更新日期：2002-12-01 00:00:00

abstract：:Tandemly polymerized regulatory elements, antisense RNA segments or ribozymes are potentially useful in selective gene silencing. However, existing methods of tandemly polymerizing short DNA segments are laborious. We present a procedure that can create cloned arrays of 40-70 monomer units in two steps. We have create...

journal_title：BioTechniques

authors： Graham GJ,Maio JJ

更新日期：1992-11-01 00:00:00

abstract：:Fluorescent dye terminator Sanger sequencing (FTSS), with detection by automated capillary electrophoresis (CE), has long been regarded as the gold standard for variant detection. However, software analysis and base-calling algorithms used to detect mutations were largely optimized for resequencing applications in whi...

journal_title：BioTechniques

authors： Davidson CJ,Zeringer E,Champion KJ,Gauthier MP,Wang F,Boonyaratanakornkit J,Jones JR,Schreiber E

更新日期：2012-09-01 00:00:00

abstract：:The colony formation assay (CFA) is the gold standard for measuring the effects of cytotoxic agents on cancer cells in vitro; however, in its traditional 6-well format, it is a time-consuming assay, particularly when evaluating combination therapies. In the interest of increased efficiency, the 6-well CFA was converte...

journal_title：BioTechniques

authors： Katz D,Ito E,Lau KS,Mocanu JD,Bastianutto C,Schimmer AD,Liu FF

更新日期：2008-02-01 00:00:00

abstract：:Autoantibodies directed against the 68-kDa (U1) ribonucleoprotein antigen are mainly found in sera of patients with mixed connective tissue disease. The corresponding cDNA was fragmented into four regions coding for the major antigenic epitopes A', B', C' and D'. All the epitopes were subcloned and expressed as fusion...

journal_title：BioTechniques

authors： Frorath B,Scanarini M,Netter HJ,Abney CC,Liedvogel B,Lakomek HJ,Northemann W

更新日期：1991-09-01 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：: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：:Genotyping fish larvae is a valuable technique for numerous fields of study. While methods for collecting DNA from early stage larvae have been published, a non-lethal, non-invasive genotyping protocol for hatchlings that is amenable to high-throughput approaches is desirable. Here, we describe a method to individuall...

journal_title：BioTechniques

authors： Espinoza GJ,Poland JM,Bremer JRA

更新日期：2017-10-01 00:00:00

abstract：:Scanning conductance microscopy investigations were carried out in air on human chromosomes fixed on pre-fabricated SiO2 surfaces with a backgate. The point of the investigation was to estimate the dielectric constant of fixed human chromosomes in order to use it for microfluidic device optimization. The phase shift c...

journal_title：BioTechniques

authors： Clausen CH,Lange JM,Jensen LB,Shah PJ,Dimaki MI,Svendsen WE

更新日期：2008-02-01 00:00:00

abstract：:Planar lipid bilayers represent a versatile platform for studying the functions of various membrane proteins as well as the development of biosensors. Despite the continuing technological progress in the fabrication of low-noise bilayer setups with mechanically and electrically stable planar bilayers, there is still a...

journal_title：BioTechniques

authors： Novák P,Gaburjáková M,Zahradník I

更新日期：2007-03-01 00:00:00

abstract：:Electrophoretic mobility shift analysis (EMSA) is a well-characterized and widely used technique for the analysis of proten-DNA interaction and the analysis of transcription factor combinatorics. Currently implemented EMSA generally involves the time-consuming use of radiolabeled DNA and polyacrylamide gel electrophor...

journal_title：BioTechniques

authors： Clark J,Shevchuk T,Swiderski PM,Dabur R,Crocitto LE,Buryanov YI,Smith SS

更新日期：2003-09-01 00:00:00

abstract：:Thymidine kinase 1 (TK1) is an enzyme involved in DNA precursor synthesis that has been used as a biomarker for prognosis and monitoring of different malignancies. In this study, we compared two immunoassays for measuring TK1 protein concentrations: the TK 210 ELISA (AroCell AB) and TK1 ELISA from Abcam. Overall, the ...

journal_title：BioTechniques

authors： Kumar JK,Holmgren S,Levedahl KH,Höglund M,Venge P,Eriksson S

更新日期：2020-06-01 00:00:00

abstract：:A streamlined protocol is described that allows high sensitivity antigen detection by Western blotting in a single day. The choice of membrane blotting matrix, as well as blocking reagents, has been optimized in order to allow rapid development of the blot with chemiluminescent reagents. The entire process, from gel t...

journal_title：BioTechniques

authors： Klapper A,MacKay B,Resh MD

更新日期：1992-05-01 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

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：: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：: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：:We describe a simple and efficient technique that facilitates the amplification of specific mRNA for cloning and sequencing purposes. An mRNA bound to a small piece of membrane filter is used as a template to synthesize complementary DNA. The product of this reaction is then transferred to a new tube and amplified usi...

journal_title：BioTechniques

authors： Ruiz A,Bok D

更新日期：1993-11-01 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：: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：:Gene expression studies require analysis of RNA, but isolation of total RNA from very small samples by traditional methods can be difficult and inefficient. The Absolutely RNA microprep kit provides a convenient method for isolating total RNA from small numbers of cells such as those harvested by laser capture microdi...

journal_title：BioTechniques

authors： Dolter KE,Braman JC

更新日期：2001-06-01 00:00:00

abstract：:A method is described for preparing mutants with multiple, site-directed mutations by ordered coupling of PCR-generated fragments catalyzed by a thermostable DNA ligase. Annealing of the sense strands of the fragments to a single-stranded (antisense) template created a full-length sense strand leaving only nicks that ...

journal_title：BioTechniques

authors： Rouwendal GJ,Wolbert EJ,Zwiers LH,Springer J

更新日期：1993-07-01 00:00:00

abstract：:An improved method was developed for in situ reverse transcription-polymerase chain reaction (RT-PCR) to detect and localize mRNA in tissue sections. The coverslip mounted-immersion cycled (COSMIC) in situ RT-PCR technique combines the advantages of solution-phase PCR with the tissue immobilization necessary for in si...

journal_title：BioTechniques

authors： Ertsey R,Scavo LM

更新日期：1998-01-01 00:00:00

abstract：:Haplotypes are useful in population genetics and medicine. Determining haplotypes in the absence of DNA samples from appropriate family members can be difficult and laborious. We have developed a rapid and reproducible method for haplotyping an individual in the absence of relatives. The method utilizes pairs of allel...

journal_title：BioTechniques

authors： Sarkar G,Sommer SS

更新日期：1991-04-01 00:00:00

abstract：:Studies on the regulation of human milk protein genes and suitable cell culture systems have been limited due to restricted availability of tissue samples from lactating women. Although mammary gland tissue has been available from mammary reduction surgery, studies on tissue-specific expression of milk protein genes r...

journal_title：BioTechniques

authors： Lindquist S,Hansson L,Hernell O,Lönnerdal B,Normark J,Strömquist M,Bergström S

更新日期：1994-10-01 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