解剖学和形态学
麻醉学
听力与言语-语言病理学
行为科学
心脏和心血管系统
细胞和组织工程学
临床神经病学
危重症监护医学
牙科,口腔外科和医学
皮肤病学
急诊医学
内分泌学和新陈代谢
肠胃学和肝脏学
老人病学和老年医学
卫生保健科学和服务
血液学
免疫学
传染病
综合和补充性医学
医学伦理学
医学信息学
医学实验室技术
医学,全科和内科
医学,法律
医学,研究和试验
神经系统科学
护理
营养学和饮食学
产科医学和妇科医学
肿瘤学
眼科学
整形外科学
耳鼻喉科学
病理学
儿科学
周围血管疾病
药理学和药剂学
生理学
基本医疗保健
精神病学
公共、环境和职业卫生
放射学,核医学和医学成像
康复学
生殖生物学
呼吸系统
风湿病学
运动科学
外科学
毒理学
热带医学
泌尿学和肾脏学
病毒学
老年医学
健康政策和服务
心理学,临床
abstract::Neuronal death can be induced by DNA-damaging agents and occurs by apoptosis involving a specific signal-transduction pathway. However, to our knowledge, methods for the quantitative determination of DNA damage in individual neurons have not yet been described. Here we optimize the single-cell gel electrophoresis (SCG...
journal_title:BioTechniques
pub_type:
doi:10.2144/99262st02
更新日期:1999-02-01 00:00:00
abstract::Reverse transcription PCR (RT-PCR) represents a sensitive and powerful tool for analyzing RNA. While it has tremendous potential for quantitative applications, a comprehensive knowledge of its technical aspects is required. Successful quantitative RT-PCR involves correction for experimental variations in individual RT...
journal_title:BioTechniques
pub_type: 杂志文章,评审
doi:10.2144/99261rv01
更新日期:1999-01-01 00:00:00
abstract::Human telomerase, a ribonucleoprotein enzyme is known to be associated with immortalized cancer cells but is absent in most normal tissues. Thus, telomerase appears to be an attractive new target for anticancer agents and an important diagnostic marker of human cancers. Here, we describe an improved telomerase assay m...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/98256cr03
更新日期:1998-12-01 00:00:00
abstract::The need for a primer hybridization step before sequencing has been eliminated using a stem-loop structure generated by PCR. The loop structure is obtained by careful design of the PCR primer or by cloning the target DNA into a dedicated vector (pRIT 28HP). After solid-phase capture of the PCR product, the loop is for...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/98255rr02
更新日期:1998-11-01 00:00:00
abstract::Changes in cardiac structure that depart from normal have generally been termed "remodeling". Assessment of ventricular remodeling at the cellular level should include measurement of myocyte dimensions. A well-established and reliable method to assess myocyte remodeling uses isolated cells and the Coulter Counter/Chan...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/98253pf02
更新日期:1998-09-01 00:00:00
abstract::As a strategy to purify recombinant murine Interleukin (IL)-18, we cloned the mature coding region of this protein into the pFLAG-1 expression system. The intent was to use the FLAG peptide "tag" as an amino terminal addition to IL-18 so that purification of this fusion protein (FLAG-IL-18) on anti-FLAG antibody affin...
journal_title:BioTechniques
pub_type:
doi:
更新日期:1998-09-01 00:00:00
abstract::Here, we present a rapid and reproducible procedure based on square-wave pulse electroporation that allows efficient penetration of synthetic oligonucleotides into intact yeast cells. This procedure was successfully used to modify the yeast genome with small amounts of oligonucleotide. ...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/98252gt04
更新日期:1998-08-01 00:00:00
abstract::Recent advances in long reverse transcription (RT)-PCR technology allow the copying of full-length coding regions of large mRNAs in one step. Using long RT-PCR, one can be certain that a given cDNA is derived from a single mRNA. In what to our knowledge is a novel application, we can isolate and characterize splice va...
journal_title:BioTechniques
pub_type:
doi:10.2144/98252st01
更新日期:1998-08-01 00:00:00
abstract::A new qualitative PCR product detection assay called competitive amplified single mutation detection by selective probe hybridization immunoassay (CASSI) was developed for genotyping the most common apolipoprotein E (apoE) polymorphisms. Single target DNA strands immobilized using biotin on streptavidin-coated micropl...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/98251st03
更新日期:1998-07-01 00:00:00
abstract::Screening methods for chemotherapeutic agents usually rely on the cytotoxic properties of the drugs. However, agents that inhibit invasion may have more efficacy and cause fewer side effects. Various cellular invasion assays have been used to evaluate these types of compounds, including the modified Boyden chamber, mo...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/98246cr04
更新日期:1998-06-01 00:00:00
abstract::Previous experiments have clearly demonstrated that microtubule dynamic instability is regulated in living cells, but the molecular mechanisms that are responsible for this regulation are not well understood. We describe two rapid, functional assays that can be used to screen cell extracts for regulators of microtubul...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/98245rr04
更新日期:1998-05-01 00:00:00
abstract::14CO2 capture is commonly used to evaluate the cellular oxidation rate of respiratory substrates. A modification of the established 14CO2-capture method was developed that enables the use of cells in adherent culture and easy analysis of multiple samples under different culture conditions. The use of commercially avai...
journal_title:BioTechniques
pub_type:
doi:10.2144/98245st04
更新日期:1998-05-01 00:00:00
abstract::The use of reverse transcription (RT) PCR for relative quantitation of gene transcripts relies on the reproducibility of the individual RT, PCR and product measurement steps. Semi-competitive RT-PCR (RT-cPCR) uses an internal competitor template in the PCR step to improve quantitation. We have surveyed the reproducibi...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/98244rr02
更新日期:1998-04-01 00:00:00
abstract::A labeled peptide nucleic acid (PNA) antisense probe was used to study the spatial distribution of triplet repeats (CTG) in human myotonic dystrophy (DM) cells by high-resolution fluorescence in situ hybridization (FISH). It was found that transcripts containing triplet repeats were present as a number of discrete foc...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/98243rr02
更新日期:1998-03-01 00:00:00
abstract::Random mutagenesis followed by an in vitro selection procedure was shown to be capable of identifying important bases of the hairpin ribozyme for cleavage of an RNA target sequence. The selection scheme enriched the RNA population for those molecules capable of efficient site-specific self-cleavage in the absence of l...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/98242st05
更新日期:1998-02-01 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
pub_type: 杂志文章
doi:10.2144/98241st02
更新日期:1998-01-01 00:00:00
abstract::We have developed a new strategy for differential screening of genes that are expressed in two or more tissues, and have used it to identify genes that are preferentially expressed in both testis and ovary. In this approach, testis-specific cDNAs were first generated using the suppression subtractive hybridization tec...
journal_title:BioTechniques
pub_type:
doi:10.2144/97236st05
更新日期:1997-12-01 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
abstract::Treatment with 2 mM CuSO4 was used to induce a Drosophila melanogaster metallothionein (Mtn) promoter that had been cloned into a recombinant baculovirus. Careful study revealed that the Mtn promoter functioned as an inducible, if somewhat "leaky" promoter within the context of baculovirus-infected cells. In the proce...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/97234rr01
更新日期:1997-10-01 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
pub_type:
doi:10.2144/97233st06
更新日期:1997-09-01 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
pub_type: 杂志文章
doi:10.2144/97232pf01
更新日期:1997-08-01 00:00:00
abstract::Automated DNA sequencing requires the intensive use of computers to handle the large amount of data taken. When a computer failure occurs and the data are no longer accessible, all the expense and effort that went into the sequencing experiment is lost. By using the data storage architecture of Macintosh computers to ...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/97226bc05
更新日期:1997-06-01 00:00:00
abstract::We have developed a high-throughput, multiplex reverse transcription PCR (RTPCR) assay that is suitable for the analysis of medium-to low-copy cellular RNA transcripts from small numbers of cells (10(4)). High throughput was attained by utilizing microplate-based RNA extraction and RTPCR protocols, followed by PCR pro...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/97226st02
更新日期:1997-06-01 00:00:00
abstract::The ZEBRA protein is a transcriptional activator that induces expression of viral lytic genes in cells harboring latent Epstein-Barr virus (EBV). In this report it is shown that a derivative of ZEBRA that cannot activate transcription (Zd) can be used to detect and characterize activation domains. Three expression vec...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/97225rr02
更新日期:1997-05-01 00:00:00
abstract::The oligonucleotide ligation assay (OLA) was adapted to the genotyping of the N-arylamine-acetyltransferase (NAT2) gene. This assay allows the use of 96-well microplates and robotic workstations for high sample throughput. We found this assay to be accurate, efficient and reliable. Another advantage for epidemiologica...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/97224st03
更新日期:1997-04-01 00:00:00
abstract::A magnetic bead-based system for DNA isolation utilizing monodisperse beads was tested with the aim of producing a general approach for PCR-ready DNA. This commercially available system was originally designed for isolating PCR-ready DNA from human whole blood. We tested diverse organisms belonging to the major groups...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/97223rr01
更新日期:1997-03-01 00:00:00
abstract::Here we demonstrate the use of a mammalian two-hybrid system to study protein-protein interactions. Like the yeast two-hybrid system, this is a genetic, in vivo assay based on the reconstitution of the function of a transcriptional activator. In this system, one protein of interest is expressed as a fusion to the Gal4...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/97222pf02
更新日期:1997-02-01 00:00:00
abstract::Rapid cycle DNA amplification was continuously monitored by three different fluorescence techniques. Fluorescence was monitored by (i) the double-strand-specific dye SYBR Green I, (ii) a decrease in fluorescein quenching by rhodamine after exonuclease cleavage of a dual-labeled hydrolysis probe and (iii) resonance ene...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/97221bi01
更新日期:1997-01-01 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::Based on the requirement of a glutathione-deficient mutant strain of Saccharomyces cerevisiae to take up external glutathione for growth on synthetic media, a simple agar diffusion test for quantitative detection of total glutathione from various sources was established. Glutathione concentrations can be reliably dete...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/96215st07
更新日期:1996-11-01 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 altered the antibiotic resistance of the reporter plasmids and the pJG4-5 activation-domain and pEG202 DNA binding-domain plasmids used in the Brent interaction trap/two-hybrid system. These plasmids were each previously ampicillin-resistant, resulting in an inefficient purification of any one plasmid from a y...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/96212st02
更新日期:1996-08-01 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::In the design of oligonucleotide sequences for targeting DNA or RNA sequences, it can be difficult to identify sequences that will hybridize only to the intended target. The term "sequence-specific" or "sequence-nonspecific" is often used to describe the interactions of an oligonucleotide with a mixture of DNA or RNA....
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/96206pf01
更新日期:1996-06-01 00:00:00
abstract::Primary human fibroblasts and a series of cell lines (A549, BNL CL.2, H225, NIH 3T3 and Rat-1) are efficiently transfected by using positively charged complexes of plasmid DNA and transferrin-polylysine or polylysine in the presence of glycerol (1 molar to 1.8 molar, depending on the cell type). An increase in gene ex...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/96205rr04
更新日期:1996-05-01 00:00:00
abstract::We describe a method using an inexpensive craft glue to routinely isolate specific areas of tissue as small as 1 mm2 from paraffin sections. The tissue may be digested to release nucleic acid suitable for PCR or reverse transcription PCR. The use of this procedure obviates the requirement for manual microdissection or...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/96205st04
更新日期:1996-05-01 00:00:00
abstract::The transcription factor GATA-1 is a zinc finger DNA-binding protein essential for the development of red blood cells. When we expressed different regions of the zinc finger domain in bacteria using an isopropyl-beta-D-thiogalactoside (IPTG) inducible system, growth of bacteria harboring the active DNA-binding domain ...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/19962004684
更新日期:1996-04-01 00:00:00
abstract::A technique that can be used to isolate vector/insert junctions from clones in vectors, such as yeast artificial chromosomes and P1s, and to sequence plasmid inserts more rapidly has been developed. A vector primer is combined with single, randomly chosen oligonucleotides in PCRs, to create pools of products. With 12-...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/19962003486
更新日期:1996-03-01 00:00:00
abstract::Subtractive hybridization has been widely used for the identification of differentially expressed genes. Here we describe a simple, sensitive strategy of subtractive hybridization that involves binding the driver poly(A)+ RNA pool to paramagnetic Dynabeads Oligo (dT)25. After hybridization with target cDNA, the molecu...
journal_title:BioTechniques
pub_type:
doi:10.2144/19962003413
更新日期:1996-03-01 00:00:00
abstract::The recently developed ultrasensitive reverse transcriptase (RT) test involving a PCR step can detect minute amounts of RT in single retroviral particles and is 10(6) times more sensitive than conventional RT assays. We have found that different DNA-dependent DNA polymerases like DNA Polymerase I from Escherichia coli...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1996-02-01 00:00:00