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：:Methods have been developed to rapidly visualize the size distribution of third complementarity-determining regions (CDR3) in immunoglobulin (Ig) and T-cell receptor (TCR) molecules. DNA fragments spanning the Ig or TCR CDR3 are generated by PCR using primers at fixed positions in the variable and constant segments. T...
journal_title：BioTechniques
pub_type：
doi：10.2144/96201st02
更新日期：1996-01-01 00:00:00
abstract：:A modification of the TRI Reagent procedure has been elaborated for isolation of RNA from polysaccharide- and proteoglycan-rich material. In the modified procedure, RNA is precipitated from the aqueous phase by the combined action of isopropanol and a high-salt concentration. Under these conditions, RNA is effectively...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1995-12-01 00:00:00
abstract：:We have modified the terminal deoxynucleotidyl transferase (TDT)-mediated dUTP-biotin nick end-labeling (TUNEL) method to permit the immunogold-silver intensification detection of apoptotic cells in tissue sections. Such sections can subsequently be processed for multi-labeling fluorescence microscopy, thus permitting...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1995-11-01 00:00:00
abstract：:The green fluorescent protein (GFP) from the jellyfish Aequorea victoria is rapidly becoming an important reporter molecule for monitoring gene expression and protein localization in vivo, in situ and in real time. GFP emits bright green light (lambda max = 509 nm) when excited with UV or blue light (lambda max = 395 ...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1995-10-01 00:00:00
abstract：:A major problem in analyzing forensic casework samples is the presence of genetic material from more than one individual in the material evidence. For instance, in sexual assault cases the evidence (vaginal swabs) usually contains a majority of vaginal epithelial cells and varying amounts of sperm cells from the perpe...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1995-09-01 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：: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
pub_type：
doi：
更新日期：1995-07-01 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
pub_type：
doi：
更新日期：1995-06-01 00:00:00
abstract：:A nonradioactive functional assay was developed to quantitate DNA binding proteins. The assay was designed to allow the use of 96-well microplates for high sample throughput. We show that the assay can measure sequence-specific DNA binding of purified proteins as well as DNA binding activity present in whole cell extr...
journal_title：BioTechniques
pub_type：
doi：
更新日期：1995-06-01 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
pub_type：
doi：
更新日期：1995-05-01 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
pub_type： 杂志文章
doi：
更新日期：1995-05-01 00:00:00
abstract：:As an improved strategy for producing functional macromolecules by in vitro evolutionary optimization, we propose an automated machine that can process up to 960 samples in parallel. It consists of a 960-well PCR machine with special sealed plastic reaction vessels and appropriate handling devices. We show that the he...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1995-04-01 00:00:00
abstract：:A new procedure is described for the generation of high-titer, helper-free retrovirus vectors employing receptor-mediated, adenovirus-augmented transfection into a standard packaging cell line. Viral titers are increased 30-fold to 100-fold in transiently (> 10(5) infectious units per mL) and stable (> 10(7) infectiou...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1995-03-01 00:00:00
abstract：:Several methods and computer programs have been developed for estimating the size of DNA fragments from gel electrophoresis. However, methods are lacking that may facilitate in optimization of gel conditions. In this article, a computer program called DNAOPT is described, which was developed to assist researchers in t...
journal_title：BioTechniques
pub_type：
doi：
更新日期：1995-02-01 00:00:00
abstract：:Plasmid DNA can be purified in 20 min using a three-step CsCl gradient in a vertical tube rotor. Ultracentrifugation with discontinuous gradients yields DNA with purity comparable to that obtained with longer isopycnic separations and with columns. ...
journal_title：BioTechniques
pub_type：
doi：
更新日期：1995-01-01 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
pub_type： 杂志文章
doi：
更新日期：1995-01-01 00:00:00
abstract：:Recombinant retroviral vectors usually encode the genes of interest in place of the viral structural genes, which must be provided in trans. These viruses are therefore defective for replication: infected cells cannot produce progeny virus. However, in some cases it may be desirable to generate virus from an infected ...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1994-12-01 00:00:00
abstract：:Human multiple tissue Western (MTW) blots are premade immunoblots prepared using proteins isolated from adult human tissue. The proteins are isolated from whole tissue homogenates under conditions designed to minimize proteolysis and to ensure maximal representation of tissue-specific proteins. Sodium dodecyl sulfate ...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1994-11-01 00:00:00
abstract：:We describe a general method for making template DNA for sequencing of PCR products. The procedure may be particularly useful for PCR products where minimal sequence information is known or as an alternative to primer walking when sequencing long PCR products. A cassette containing the hybridization site for the M13 s...
journal_title：BioTechniques
pub_type：
doi：
更新日期：1994-11-01 00:00:00
abstract：:Exposing cells to brief, high-intensity electrical field pulses can lead to the permeabilization of their plasma membranes. This electro-induced permeated state of the cell membrane is reversible and leads to cell fusion; i.e., the electropermeabilized state if fusogenic. The size of cells intended for fusing, however...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1994-10-01 00:00:00
abstract：:The construction and the testing of two lambda phage vectors are described that greatly simplify the tasks of mapping genomic DNA and making replacement-type gene-targeting vectors for mammalian cells from a library of isogenic genomic DNA. The first vector, lambda PS, accommodates up to 20 kb and allows inserts to be...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1994-10-01 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
pub_type：
doi：
更新日期：1994-10-01 00:00:00
abstract：:Here we describe our progress in the development of the technology of DNA sequencing by primer walking based on the modular primer approach, which eliminates the primer synthesis bottleneck. Modular primers are assembled from 5-mers, 6-mers, or 7-mers selected from a presynthesized library of as few as 1000 oligonucle...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1994-09-01 00:00:00
abstract：:Data are presented illustrating the optimum concentration range of reverse transcription PCR-generated products under 500 bp for accurate base calling with direct automated DNA sequencing. A 357-bp fragment of the human estrogen receptor, which includes the DNA binding domain of the protein, was used as a representati...
journal_title：BioTechniques
pub_type：
doi：
更新日期：1994-09-01 00:00:00
abstract：:We describe here a simple and efficient transfection method for transient expression of cloned genes in cell lines and primary cultured cells. The method involves the use of DEAE-dextran to target DNA to the cellular endocytotic pathway and the use of a human adenovirus to ensure efficient lysis of endosomal vesicles....
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1994-08-01 00:00:00
abstract：:A simple spectrophotometric method for measuring DNA in proliferating cells is described. The method is an adaptation of the widely used diphenylamine (DPA) reaction to examine DNA in cells grown in a 96-well plate. This assay was capable of detecting as little as 10 ng DNA and could be used to measure DNA in stored a...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1994-07-01 00:00:00
abstract：:A brief description of the setup of an inexpensive densitometric analysis system is provided. This system uses a software package and scanner that can be obtained for a total of less than $1000 and that can be operated by any Macintosh computer, including a MacPlus. In the event that a scanner is already available, th...
journal_title：BioTechniques
pub_type：
doi：
更新日期：1994-06-01 00:00:00
abstract：:A PCR-based method is described for the production of cDNA libraries and total cDNA probes from a few milligrams of tissue. Using a model system, we show how a PCR library and PCR probes can be used to identify genes expressed at different levels in two tissues. Small amounts of tissue derived from two plants, one inf...
journal_title：BioTechniques
pub_type：
doi：
更新日期：1994-06-01 00:00:00
abstract：:We have developed a procedure for undertaking a Microtox-based test by coupling microplate and microluminometric technologies. Sample dilutions are prepared in a 96-well polystyrene microplate kept at 15 degrees C, while the Microtox reagent and diluent are placed in an opaque, microluminometry-compatible 96-well micr...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1994-05-01 00:00:00
abstract：:DNA-binding proteins can be purified by their affinity for probes with specific sequences that are tethered to magnetic beads. A restriction site at the end of the tether allows release of probe with factor still on it. This probe with bound factor can be bandshifted directly to compare to bandshifts in whole extracts...
journal_title：BioTechniques
pub_type：
doi：
更新日期：1994-05-01 00:00:00
abstract：:Much effort has been directed toward the isolation and characterization of homeobox cDNAs from numerous cell types because they encode transcription factors important to many cellular processes, including pattern formation in the embryo, cell growth and cell differentiation. Many novel homeobox cDNAs have been isolate...
journal_title：BioTechniques
pub_type：
doi：
更新日期：1994-05-01 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
pub_type：
doi：
更新日期：1994-04-01 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：:A novel method for large-scale plasmid preparation is described. Crude extracts are subjected to acidic phenol extraction to remove any contaminants present in the aqueous phase. The supercoiled plasmid DNA, which preferentially remains in the organic phase and inter-phase, is extracted back into the aqueous phase wit...
journal_title：BioTechniques
pub_type：
doi：
更新日期：1994-03-01 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：: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
pub_type：
doi：
更新日期：1994-01-01 00:00:00
abstract：:The Sequence Analysis Workshop (SAW) is an interactive program for sequence analysis of immunoglobulin variable domains. Sequences for SAW can be obtained from GenBank or from a standard text file. SAW can compare a variable domain to as many as 100 different sequences, calculate the extent of homology, sort the seque...
journal_title：BioTechniques
pub_type：
doi：
更新日期：1993-12-01 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
pub_type：
doi：
更新日期：1993-11-01 00:00:00
abstract：:The serum protein transferrin (Tf) is a valuable marker for genetic studies of primates, because it is usually polymorphic, exhibiting as many as 13 allelic forms with high heterozygosity. The standard procedure to detect the different phenotypes requires vertical electrophoresis on polyacrylamide gels for 18 h at 4 d...
journal_title：BioTechniques
pub_type：
doi：
更新日期：1993-11-01 00:00:00
abstract：:The purpose of this research was to compare three bioreactor systems for the pilot-scale production of monoclonal antibodies (MAbs). We needed to produce gram quantities of murine MAbs against human prostatic acid phosphatase for use in fragmentation, radiolabeling and in vivo radio-imaging studies. The stable hybrido...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1993-10-01 00:00:00
