Antibiotic-free bacterial strain selection using antisense peptide nucleic acid.

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：: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：:The two-hybrid system is a genetic method to identify proteins that interact with a specific target protein, which is expressed in yeast as a hybrid with a DNA-binding domain. Use of this method entails transforming yeast, both with this DNA-binding domain hybrid and with a library of activation domain hybrids, follow...

journal_title：BioTechniques

authors： Bartel P,Chien CT,Sternglanz R,Fields S

更新日期：1993-06-01 00:00:00

abstract：:Virus uptake is the first rate-limiting step for the successful gene delivery of any virus-based gene therapy. For adenovirus-based gene therapy, the expression levels of the adenovirus receptor--coxsackievirus and adenovirus receptor (CAR)--play an important role in dictating gene delivery. We have observed a wide sp...

journal_title：BioTechniques

authors： Lai YJ,Pong RC,McConnell JD,Hsieh JT

更新日期：2003-07-01 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：:Display of functional antibody fragments on the surface of filamentous bacteriophages allows fast selection of specific phage antibodies against a variety of target antigens. However, enrichment of single chain variable fragment (scFv)-displaying phages is often hampered by the abundance of bacteriophages lacking anti...

journal_title：BioTechniques

authors： Tur MK,Huhn M,Sasse S,Engert A,Barth S

更新日期：2001-02-01 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 consensus peptide sequence, QSYP, appears as an artifact during the mapping of monoclonal antibodies (MAbs) using a random peptide phage display library. Phage bearing this QSYP sequence were independently selected by four different laboratories screening separate MAb preparations with the same phage library. In eac...

journal_title：BioTechniques

authors： Jacobs JM,Bailey BW,Burritt JB,Morrison SG,Morrison RP,Dratz EA,Jesaitis AJ,Teintze M

更新日期：2003-01-01 00:00:00

abstract：:DNA damage in the form of abasic sites, chemically altered nucleotides, and strand fragmentation is the foremost limitation in obtaining genetic information from many ancient samples. Upon cell death, DNA continues to endure various chemical attacks such as hydrolysis and oxidation, but repair pathways found in vivo n...

journal_title：BioTechniques

authors： Mouttham N,Klunk J,Kuch M,Fourney R,Poinar H

更新日期：2015-07-01 00:00:00

abstract：:We describe a simple and cost-effective method for the synthesis of an internal fluorescently labeled DNA standard for fragment sizing using an automated DNA sequencer. A set of primer pairs labeled with ROX was developed to amplify 12 DNA fragments, 58-417 bp, derived from a conserved region of plant chloroplast DNA....

journal_title：BioTechniques

authors： Brondani RP,Grattapaglia D

更新日期：2001-10-01 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：:Current gene synthesis methods often incorporate a PCR amplification step in order to yield final material sufficient for resolution from multiple off-products. These amplification steps can cause stochastic sampling effects that propagate errors in gene synthesis or decrease variability when applied to the constructi...

journal_title：BioTechniques

authors： Lee ZB,Firnhaber C,Clarke J,DeDecker BS

更新日期：2015-09-01 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：: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：:To increase the reproducibility and to reduce the false positives in the initial mRNA differential display, modified long composite primers were developed based on both mRNA differential display and RNA arbitrarily primed PCR fingerprinting methods. Ten-base nucleotides were added at the 5' ends of the primers used in...

journal_title：BioTechniques

authors： Zhao S,Ooi SL,Pardee AB

更新日期：1995-05-01 00:00:00

abstract：:Transfection of suspension cells has proven to be very difficult using conventional methods. Here, we present a simple and time-saving new transfection protocol wherein cell culture plates coated with chicken egg white are seeded with suspension cells prior to transfection. Our results demonstrate that coupling egg wh...

journal_title：BioTechniques

authors： Basiouni S,Fuhrmann H,Schumann J

更新日期：2012-08-01 00:00:00

abstract：:A 200-year-old observation might provide a new way to eliminate tumors by infecting cancer patients with bacteria. Kristie Nybo explores a new approach that could transform cancer treatment. ...

journal_title：BioTechniques

authors： Nybo K

更新日期：2018-01-01 00:00:00

abstract：:The technique described here is a fast and simple method of extracting chloroplast DNA (cpDNA). It overcomes the need for differential centrifugation using density gradients. The leaves do not have to be kept in the dark and lyophilized before extraction, but lyophilization is still possible. The chloroplasts are spec...

journal_title：BioTechniques

authors： Mariac C,Trouslot P,Poteaux C,Bezançon G,Renno JF

更新日期：2000-01-01 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：:Microarray expression analysis has become one of the most widely used functional genomics tools. Efficient application of this technique requires the development of robust and reproducible protocols. We have optimized all aspects of the process, including PCR amplification of target cDNA clones, microarray printing, p...

journal_title：BioTechniques

authors： Hegde P,Qi R,Abernathy K,Gay C,Dharap S,Gaspard R,Hughes JE,Snesrud E,Lee N,Quackenbush J

更新日期：2000-09-01 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：: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：:Three-dimensional collagen gel contraction is the standard assay utilized for functionally quantifying a variety of cell types, in particular smooth muscle cells (SMCs) and myofibroblasts. Here, we have developed a method to effectively reduce the three-dimensional parameters of the standard collagen gel into a single...

journal_title：BioTechniques

authors： Ilagan R,Guthrie K,Quinlan S,Rapoport HS,Jones S,Church A,Basu J,Ludlow J

更新日期：2010-02-01 00:00:00

abstract：:The increase of information in biology makes it difficult for researchers in any field to keep current with the literature. The MEDLINE database of scientific abstracts can be quickly scanned using electronic mechanisms. Potentially interesting abstracts can be selected by matching words joined by Boolean operators. H...

journal_title：BioTechniques

authors： Perez-Iratxeta C,Keer HS,Bork P,Andrade MA

更新日期：2002-06-01 00:00:00

abstract：:3D printed biomaterials are increasingly used in cell cultures and drug screens. Given the ease of creating artificial tissues, will this technique revolutionize biomedicine and organ implants in the future? ...

journal_title：BioTechniques

authors： Prabhune M

更新日期：2018-03-01 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：:Replication studies on human immunodeficiency virus 1 (HIV-1) rely on a few laboratory strains that are divergent from dominant HIV-1 subtypes in the epidemic. Several phenotypic differences between diverse HIV-1 isolates and subtypes could affect vaccine development and treatment, but this research field lacks robust...

journal_title：BioTechniques

authors： Dudley DM,Gao Y,Nelson KN,Henry KR,Nankya I,Gibson RM,Arts EJ

更新日期：2009-05-01 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：:GC-rich DNA regions were PCR-amplified with Taq DNA polymerase using either the canonical set of deoxynucleoside triphosphates or mixtures in which the dCTP had been partially or completely replaced by its N4-methylated analog, N4-methyl-2'-deoxycytidine 5'-triphosphate (N4me-dCTP). In the case of a particularly GC-ri...

journal_title：BioTechniques

authors： Flores-Juárez CR,González-Jasso E,Antaramian A,Pless RC

更新日期：2016-10-01 00:00:00

abstract：:The possibility to miniaturize and parallelize biological assays has a great impact on the development of biomedical technologies. Here, we describe a simple, miniaturized, and parallelized method employing entire cells from different cell lines displaying a protein of interest on their surface, which were immobilized...

journal_title：BioTechniques

authors： Schwenk JM,Stoll D,Templin MF,Joos TO

更新日期：2002-12-01 00:00:00