Use of perfluorocarbon emulsions in cell culture.

abstract：:RNA interference (RNAi) is a recent advance that provides the possibility to reduce the expression of specific target genes in cultured mammalian cells with potential applications on a genome-wide scale. However, to achieve this, robust methodologies that allow automated and efficient delivery of small interfering RNA...

journal_title：BioTechniques

authors： Erfle H,Simpson JC,Bastiaens PI,Pepperkok R

更新日期：2004-09-01 00:00:00

abstract：:To avoid the time-consuming reprobing process in hybridization analysis, signal-distinguishable probes (32P, 35S or antigenic hapten-labeled DNA) can be added to the same hybridization mixture. After hybridization, an unambiguous result can be obtained by differential or sequential autoradiography. ...

journal_title：BioTechniques

authors： Au LC,Chang KJ,Shih CM,Teh GW

更新日期：1994-04-01 00:00:00

abstract：:New cloning vectors have been developed with features to enhance quick allelic exchange in gram-negative bacteria. The conditionally replicative R6K and transfer origins facilitate conjugation and chromosomal integration into a variety of bacterial species, whereas the sacB gene provides counterselection for allelic e...

journal_title：BioTechniques

authors： Mejia-Santana A,Lloyd CJ,Klose KE

更新日期：2021-01-25 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：:Antibiotics are widely useful in medicine, agriculture, and industrial fermentations. However, increasing problems with resistant strains call for restrained use and alternative strategies. Antisense peptide nucleic acids (PNAs) show potent bactericidal effects when targeted against the essential Escherichia coli acpP...

journal_title：BioTechniques

authors： Dryselius R,Nekhotiaeva N,Nielsen PE,Good L

更新日期：2003-11-01 00:00:00

abstract：:Gel retardation analysis, or band shift assay, is technically the simplest method to investigate protein-nucleic acid interactions. In this report, we describe a nonradioactive band shift assay using a fluorescent DNA target and an ALFexpress automatic DNA sequencer in place of the current method that utilizes radioac...

journal_title：BioTechniques

authors： Filée P,Delmarcelle M,Thamm I,Joris B

更新日期：2001-05-01 00:00:00

abstract：:Immunofluorescence quantification of γH2AX foci is a powerful approach to quantify DNA double-strand breaks induced by cancer therapy or accidental exposure to ionizing radiation. Here we report a modification to the γH2AX immunofluorescence labeling method, whereby cells are stained in-solution before being spotted a...

journal_title：BioTechniques

authors： Johansson P,Muslimovic A,Hultborn R,Fernström E,Hammarsten O

更新日期：2011-09-01 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：:This report describes a common method of obtaining template DNA from phagemids, phages and plasmids. The strategy is based on the use of the cationic detergent cetyl-trimethylammonium bromide (CTAB) for DNA precipitation. By avoiding phase separation, many manipulation steps are reduced. A time-saving modification to ...

journal_title：BioTechniques

authors： Del Sal G,Manfioletti G,Schneider C

更新日期：1989-05-01 00:00:00

abstract：:There is an enormous initiative to establish the genetic basis for disorders of brain function. Unfortunately, genetic intervention is not accomplished easily in the nervous system. One strategy is to engineer and deliver to neurons specialized viral vectors that carry a gene (or genes) of interest, thereby exploiting...

journal_title：BioTechniques

authors： Neve RL,Neve KA,Nestler EJ,Carlezon WA Jr

更新日期：2005-09-01 00:00:00

abstract：:To gain insightful information about the mechanisms through which genes are activated and repressed requires gene reporter systems that are sensitive, robust, and cost-effective. Although numerous reporter gene technologies are commercially available, none are as sophisticated and user-friendly as beta-lactamase (BLA)...

journal_title：BioTechniques

authors： Qureshi SA

更新日期：2007-01-01 00:00:00

abstract：:Microarray technologies have made possible comprehensive analyses of nucleic acid sequence and expression. However, the technology to obtain efficiently high-quality RNA and DNA suitable for array analysis from purified populations of neoplastic cells from human tissues has not been well addressed. Microdissection can...

journal_title：BioTechniques

authors： Barrett MT,Glogovac J,Prevo LJ,Reid BJ,Porter P,Rabinovitch PS

更新日期：2002-04-01 00:00:00

abstract：:Polyethyleneimine (PEI) is a flocculent that is widely used in the downstream purification of monoclonal antibodies. It is an in-process residual that is carried through the drug purification process and strongly inhibits residual DNA quantitation by real-time quantitative PCR assay. Very high sample dilutions (e.g., ...

journal_title：BioTechniques

authors： Zhang SM,Roberts M,Jones M,Zeitler J,Kilby G,Liu A,White JR

更新日期：2020-06-01 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：:Quantitative PCR and reverse transcription PCR (RT-PCR) are widely used in biomedical, industrial and other research applications to determine the number of RNA or DNA molecules of a specific type and/or sequence in a sample of interest. We have developed an assay system to accurately quantitate PCR products that util...

journal_title：BioTechniques

authors： Martin CS,Butler L,Bronstein I

更新日期：1995-05-01 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：:Piezoelectric dispensing of proteins from borosilicate glass capillaries is a popular method of protein biochip fabrication that offers the advantages of sample recovery and noncontact with the printing substrate. However, little regard has been given to the quantitative aspects of dispensing minute volumes (1 nL or l...

journal_title：BioTechniques

authors： Delehanty JB,Ligler FS

更新日期：2003-02-01 00:00:00

abstract：:Selective delivery of genes to ocular tissues in vivo has been a long sought after goal for potential gene therapy of ocular disease. The gene gun was considered for this purpose because of its ability to focally transfer DNA to cells through gold microparticles coated with DNA. Through experimentation, we optimized a...

journal_title：BioTechniques

authors： Tanelian DL,Barry MA,Johnston SA,Le T,Smith G

更新日期：1997-09-01 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：:We describe a method for rapid radioactive labeling of PCR product. The method, employing the Klenow fragment of DNA polymerase I, consumes little product, requires no product purification and takes under 30 minutes. ...

journal_title：BioTechniques

authors： McDaniel TK,Huang Y,Yin J,Needleman SW,Meltzer SJ

更新日期：1991-08-01 00:00:00

abstract：:A method for conserving primers and differently labeled fluorogenic hydrolysis (i.e., TaqMan) probes at ambient conditions is presented. Primers and hydrolysis probes with four different fluorophore-quencher combinations (6- FAM-BHQ1, HEX-BHQ1, ROX-BHQ650, and Cy5-BHQ2) were mixed with trehalose and xanthan at final c...

journal_title：BioTechniques

authors： Rombach M,Kosse D,Faltin B,Wadle S,Roth G,Zengerle R,von Stetten F

更新日期：2014-09-01 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：: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：: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：:The use of quantitative PCR is recommended to monitor the level of residual hematological malignancies. The proposed multiplex IgH/ras PCR uses a co-amplification of the clonal CDR3 rearrangement of the immunoglobulin heavy chain gene (IgH) as a disease marker and a segment of the Hras 1 gene containing codon 61 (ras)...

journal_title：BioTechniques

authors： Slavícková A,Ullmannová V,Klener P

更新日期：2000-04-01 00:00:00

abstract：:We describe a procedure for delivery of purified proteins into a variety of tissue culture cells using a new polycationic lipid preparation, LipofectAMINE. Several different proteins, with diverse physical properties, can be delivered into cells by this method. Compared with commercially available monocationic lipids,...

journal_title：BioTechniques

authors： Sells MA,Li J,Chernoff J

更新日期：1995-07-01 00:00:00

abstract：:Sensitive nucleic acid based detection methods such as in situ PCR, in situ RT-PCR and PRINS have great potential in the areas of developmental biology, pathogenesis and diagnostics. However, control of evaporation from in situ reactions is critical to ensure reliable data. Self-Seal Reagent, a component added directl...

journal_title：BioTechniques

authors： Sullivan DE,Bobroski LE,Bagasra O,Finney M

更新日期：1997-08-01 00:00:00

abstract：:A complete image digitizing and processing system is described for capturing, enhancing and analyzing molecular fingerprints. The low-cost, high-resolution system features a Motorola 68000 processor, multi-tasking, a separate video coprocessor, and color or gray scale processing. Thousands of manipulations are possibl...

journal_title：BioTechniques

authors： Mitchell BW,Palmieri S

更新日期：1990-10-01 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：: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