Abstract:
:A new method has been developed to purify and detritylate milligram amounts of synthetic oligonucleotides. Dimethoxytrityl oligonucleotides from 15 to 100 nucleotides in length are applied in triethylammonium acetate or concentrated ammonium hydroxide to a disposable chromatographic cartridge, the NENSORB PREP Nucleic Acid Purification Cartridge. Salts, failure sequences and synthetic by-products are washed away while the desired, full-length, dimethoxytrityl oligonucleotide remains bound to the cartridge. The trityl group is hydrolyzed from the 5'-end of the oligonucleotide with an acid wash and then the purified oligonucleotide is eluted with 35% methanol. Oligonucleotides are recovered salt-free with purities greater than 95%. NENSORB PREP-purified primers provide superior sequence data compared to similar primers used without purification and equivalent data to primers purified by polyacrylamide gel electrophoresis when used in manual radiometric Sanger sequencing.
journal_name
Biotechniquesjournal_title
BioTechniquesauthors
Johnson BA,McClain SG,Doran ER,Tice G,Kirsch MAsubject
Has Abstractpub_date
1990-04-01 00:00:00pages
424-9issue
4eissn
0736-6205issn
1940-9818journal_volume
8pub_type
杂志文章相关文献
BIOTECHNIQUES文献大全abstract::The microcomputer-based image analysis system IB-1000 (developed by Indiana Biotech, Highland, IN) for two-dimensional electrophoresis gels has been described previously (9). It allows the user to compare protein spots between two profiles and identify those spots that are commonly shared in both profiles. This report...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1989-04-01 00:00:00
abstract::SNPCEQer identifies and reports SNPs in sequences obtained from the Beckman CEQ2000 DNA Analysis System. SNPCEQer aligns sequences obtained using CEQ2000 heterozygote detection analysis and reports discrepancies between individual sequences and the consensus sequence it generates from this set as SNPs when the individ...
journal_title:BioTechniques
pub_type:
doi:10.2144/02334st05
更新日期:2002-10-01 00:00:00
abstract::To move microarray technology into the diagnostic realm, the impact of technical parameters, such as sample preparation and RNA extraction, needs to be understood and minimized. We evaluated the impact of two RNA extraction methods, DNase treatment and the amount of hybridized cDNA probe, on the outcome of microarray ...
journal_title:BioTechniques
pub_type:
doi:10.2144/03355st04
更新日期:2003-11-01 00:00:00
abstract::[Formula: see text] Known to be a sturdy weapon in a scientist's arsenal, how has the gene editing tool CRISPR been applied in the fight against COVID-19? ...
journal_title:BioTechniques
pub_type: 新闻
doi:10.2144/btn-2020-0145
更新日期:2020-11-01 00:00:00
abstract::Cysts of free-living protozoa have an impact on the ecology and epidemiology of bacteria because they may act as a transmission vector or shelter the bacteria against hash environmental conditions. Detection and localization of intracystic bacteria and examination of the en- and excystment dynamics is a major challeng...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000114274
更新日期:2015-04-01 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
pub_type:
doi:
更新日期:1994-02-01 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
pub_type: 杂志文章
doi:10.2144/000112757
更新日期:2008-02-01 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
pub_type: 杂志文章
doi:
更新日期:2003-11-01 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
pub_type:
doi:10.2144/01304st03
更新日期:2001-04-01 00:00:00
abstract::Single-stranded long oligonucleotide-based (50- to 70-mer) microarrays offer several advantages over conventional cDNA microarrays. These include the easy preparation of the probes, low cost of array production, and low cross-contamination during probe handling. However, the application of oligonucleotide microarrays ...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/04374ST02
更新日期:2004-10-01 00:00:00
abstract::Oligonucleotide primers used to amplify target DNA regions via PCR should meet certain design criteria to maximize the potential for efficient priming. The Random Oligonucleotide Construction Kit (ROCK), a spreadsheet-based program that runs under Microsoft Excel 97 or later version for Microsoft Windows, was develope...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/01306bc01
更新日期:2001-06-01 00:00:00
abstract::Recombinant alphaviruses have been used as vehicles for delivery and expression of heterologous genes in mammalian, avian and insect cell lines. We have used a Sindbis replicon virus (Sinreplac) able to express the E. coli lacZ gene to compare the efficiency of transduction in one insect, six mammalian cell lines and ...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/96213rr02
更新日期:1996-09-01 00:00:00
abstract::We have designed and built a magnetic tweezers device that enables the application of calibrated stresses to soft materials while simultaneously measuring their microscale deformation using confocal microscopy. Unlike previous magnetic tweezers designs, our device is entirely portable, allowing easy use on microscopes...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000113701
更新日期:2011-07-01 00:00:00
abstract::A rapid and inexpensive method for isolating bacterial DNA for use in PCR is described. The method is based on the guanidinium thiocyanate (GuSCN)-lysis method of Boom et al. (J. Clin. Microbiol. 28:495-503, 1990) and enables a multiple of 96 samples to be prepared in only one hour. We use Multiscreen plates and a vac...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1995-08-01 00:00:00
abstract::Genetic immunization is a simple method for producing polyclonal antibodies in mice. To test if this approach could be used for monoclonal antibody production, biolistic transfection was used to immunize a mouse. High levels of polyclonal antibodies against human growth hormone (hGH) were elicited following three inoc...
journal_title:BioTechniques
pub_type:
doi:
更新日期:1994-04-01 00:00:00
abstract::For a feasible and cost-effective impedance measurement of cellular alterations in real-time, we combined commercially available microelectrode arrays (MEAs), consisting of 60 microelectrodes, with a conventional impedance analyzer. For proof of principle, a breast carcinoma cell line (MCF-7) was cultured on MEAs, and...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000112254
更新日期:2006-10-01 00:00:00
abstract::There are little independent data available about how well single nucleotide polymorphism (SNP) genotyping technologies perform in the typical molecular genetics laboratory. We evaluated the utility and accuracy of a widely used technology, template-directed dye-terminator incorporation with fluorescence-polarization ...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/02325rr02
更新日期:2002-05-01 00:00:00
abstract::The role model systems have played in understanding telomere biology has been enormous, and understanding has rapidly transferred to human telomere research. Most work using model organisms to study telomerase and nontelomerase-based telomere-maintenance systems has centered on yeasts, ciliates, and insects. But it is...
journal_title:BioTechniques
pub_type: 杂志文章,评审
doi:10.2144/05382RV01
更新日期:2005-02-01 00:00:00
abstract::Metabolic and electrical coupling through gap junction channels is implicated in cell differentiation, tissue homeostasis, and electrotonic propagation of signals in excitable tissues. The characterization of gating properties of these channels requires electrophysiological recordings at both single- and multiple-chan...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/03345rr03
更新日期:2003-05-01 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
pub_type: 杂志文章
doi:10.2144/000114177
更新日期:2014-06-01 00:00:00
abstract::Naturstoff reagent A (diphenylboric acid 2-aminoethyl ester [DPBA]) has been used historically in plant science to observe polyphenolic pigments, such as flavonoids, whose fluorescence requires enhancement to be visible by microscopy. Flavonoids are common dietary constituents and are the focus of considerable attenti...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/btn-2018-0084
更新日期:2019-02-01 00:00:00
abstract::We have utilized 5-bromo-2'deoxyuridine (BrdU) substituted DNA as a probe for a number of applications including, principally, for chromosome painting by fluorescence in situ hybridization (FISH) but also for DNA end-labeling to detect apoptotic cell death and for filter hybridization. Br-dUTP was used as a substitute...
journal_title:BioTechniques
pub_type:
doi:
更新日期:1996-07-01 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
pub_type: 杂志文章
doi:
更新日期:1991-01-01 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
pub_type: 杂志文章
doi:10.2144/96216bc01
更新日期:1996-12-01 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
pub_type: 杂志文章
doi:
更新日期:1991-06-01 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
pub_type:
doi:10.2144/02336bc03
更新日期:2002-12-01 00:00:00
abstract::Mutations in the human MYH7 gene, encoding a slow skeletal muscle/β-cardiac myosin heavy chain, cause different types of myopathies. The nematode model Caenorhabditis elegans has frequently been employed to study the molecular and physiological consequences of MYH7 mutations in muscle function by introducing mutations...
journal_title:BioTechniques
pub_type: 信件
doi:10.2144/btn-2020-0012
更新日期:2020-06-01 00:00:00
abstract::Another method has been developed to attach synthetic peptides to solid supports for use in enzyme immunoassays. The method is based on passively adsorbing a synthetic peptide to a solid-phase support, then further attaching more of the same peptide by means of cross-linking to the previously adsorbed peptide. This me...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1992-11-01 00:00:00
abstract::S-palmitoylation (S-acylation) is emerging as an important dynamic post-translational modification of cysteine residues within proteins. Current assays for protein S-palmitoylation involve either in vivo labeling or chemical cleavage of S-palmitoyl groups to reveal a free cysteine sulfhydryl that can be subsequently l...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000114516
更新日期:2017-02-01 00:00:00
abstract::Ribonucleases H (RNases H) are enzymes that specifically degrade the RNA of RNA-DNA hybrids. These enzymes are involved in DNA replication, reverse transcription (RT) and antisense oligodeoxyribonucleotide-mediated arrest of translation. One of the most valuable tools for assaying RNase H activity is the renaturation ...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/97235rr01
更新日期:1997-11-01 00:00:00