Abstract：
:Current protocols for screening proliferative factors for β cells ex vivo are time-consuming, require cell lines or dissociated islets, and often entail expensive specialized screening equipment. Here we present an efficient and lower cost alternative that utilizes intact mouse islets for the initial screening of proliferative compounds and allows confirmation of β cell proliferation from the same islets. This protocol simplifies the process, saves money, and provides a way to identify β cell proliferative factors using equipment that is ubiquitous in most laboratories.
journal_name
Biotechniquesjournal_title
BioTechniquesauthors
Mosser RE,Gannon Mdoi
10.2144/000114115subject
Has Abstractpub_date
2013-12-01 00:00:00pages
310-2issue
6eissn
0736-6205issn
1940-9818pii
000114115journal_volume
55pub_type
杂志文章相关文献
BIOTECHNIQUES文献大全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：:Transfection experiments involving mixed cell cultures pose special problems because of differential uptake and expression of DNA by different cell types. In order to evaluate the expression of transfected DNA in primary cultures of embryonic myocardial cells, we have used immunofluorescence microscopy to analyze expr...
journal_title：BioTechniques
pub_type： 杂志文章,评审
doi：
更新日期：1988-07-01 00:00:00
abstract：:Gene expression analysis using high-density cDNA or oligonucleotide arrays is a rapidly emerging tool for transcriptomics, the analysis of the transcriptional state of a cell or organ. One of the limitations of current methodologies is the requirement of a relatively large amount of total or polyadenylated RNA as star...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/03343rr01
更新日期：2003-03-01 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
pub_type： 新闻
doi：10.2144/btn-2017-0113
更新日期：2018-03-01 00:00:00
abstract：:Patents play an increasingly important role in the dissemination of information in many fast moving fields such as biotechnology and semiconductors. Quite a few new developments are introduced as patents, and only later, if at all, do they find their way into the scientific literature. In spite of this, patents lack w...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1988-02-01 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
pub_type： 杂志文章
doi：
更新日期：2002-12-01 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
pub_type： 杂志文章,评审
doi：10.2144/000112292
更新日期：2007-01-01 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：:Effective data analysis of the modern biological microscopy data set often necessitates a variety of different analysis strategies, and this often means the biologist may need to use a combination of software tools both commercial and often-times open source. To facilitate this process, there needs to be knowledge of ...
journal_title：BioTechniques
pub_type： 杂志文章,评审
doi：10.2144/000112511
更新日期：2007-07-01 00:00:00
abstract：:A direct method to study essential genes is to construct conditional knock-down mutants by replacement of their native promoter by an inducible one. In Mycobacteria, replacement of an essential gene promoter with an anhydrotetracycline inducible one was successfully used but required a multi-step approach. In this wor...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/btn-2018-0074
更新日期：2018-09-01 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：:We have compared RNA polymerase promoter activities in PCR-generated DNA fragments for use in the in vitro transcription of cRNA probes. Sense oligonucleotide primers, specific for the mouse acidic fibroblast growth factor gene, were synthesized with 5' extensions containing promoter sequences for the T7, T3 and SP6 R...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1992-10-01 00:00:00
abstract：:Normalization is a critical step in the analysis of microarray gene expression data. For dual-labeled array, traditional normalization methods assume that the majority of genes are non-differentially expressed and that the number of overexpressed genes approximately equals the number of under-expressed genes. However,...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/000113195
更新日期：2009-08-01 00:00:00
abstract：:Here we describe a convenient method to generate homologous recombinant baculoviral genomes in E. coli. The recombination takes place with the aid of recombination enzymes provided by the phage lambda Red system between a bacmid (a baculoviral genome that can replicate in bacteria) and a linear fragment. Proof of conc...
journal_title：BioTechniques
pub_type：
doi：10.2144/02324st04
更新日期：2002-04-01 00:00:00
abstract：:Specific amplification of nucleic acid sequences by PCR has been extensively used for the detection of gene rearrangements and gene expression. Although successful amplification of DNA sequences has been carried out with DNA prepared from formalin-fixed, paraffin-embedded (FFPE) tissues, there are only a few reports r...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1993-03-01 00:00:00
abstract：:Here we describe an amplification method for global transcript analysis. The strategy relies on amplification of cDNA tags (signature tags) achieved by random fragmentation of the cDNAs to short tags of similar length, isolation of the 3' ends and then PCR amplification of the 3'-end signature tag population. This met...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/04362ST02
更新日期：2004-02-01 00:00:00
abstract：:We have developed an improved subtractive hybridization method that provides a fast, simple and reliable isolation of desired different sequences from two compared DNA libraries, one of which contains all unwanted homologues (subtracter) and another contains certain desired heterologues (tester). The DNA library can b...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/99265rr06
更新日期：1999-05-01 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
pub_type： 杂志文章
doi：10.2144/01305rr03
更新日期：2001-05-01 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
pub_type： 杂志文章
doi：10.2144/99272st04
更新日期：1999-08-01 00:00:00
abstract：:Methylation of DNA in CpG dense regions of gene promoters (CpG islands) is important for transcriptional inactivation of selective genes in normal and neoplastic cells. Here, we present a spreadsheet-based program adapted from Microsoft Excel that is useful for identifying CpG islands and for assisting in the laborato...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/01301bc01
更新日期：2001-01-01 00:00:00
abstract：:A streamlined protocol is described that allows high sensitivity antigen detection by Western blotting in a single day. The choice of membrane blotting matrix, as well as blocking reagents, has been optimized in order to allow rapid development of the blot with chemiluminescent reagents. The entire process, from gel t...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1992-05-01 00:00:00
abstract：:We describe here a simple and rapid method for enzymatic DNA amplification using DNA template recovered from membrane filters previously used in hybridization analysis. This is done by first solubilizing membrane pieces carrying DNA of interest in dimethyl sulfoxide, followed by isopropanol precipitation and polymeras...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1993-04-01 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：:Antibodies specific to avian myeloblastosis virus envelope glycoprotein gp80 were raised. Immunoliposomes were prepared using anti-avian myeloblastosis virus envelope glycoprotein gp80 antibody. The antibody was palmitoylated to facilitate its incorporation into lipid bilayers of liposomes. The fluorescence emission s...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1991-08-01 00:00:00
abstract：:The expression of foreign proteins in Saccharomyces cerevisiae is a powerful tool for basic research and the biotechnological industry. In spite of the potential of S. cerevisiae, only a few useful expression vectors have been developed for this yeast. These vectors are based on an increasing transcription rate in com...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/00283rr04
更新日期：2000-03-01 00:00:00
abstract：:We describe a rapid, simple and inexpensive method for the isolation of DNA from blood clots suited for use in PCR. Our method is based on the lysing and nuclease-inactivating properties of guanidine thiocyanate together with the nucleic acid-binding properties of silica particles. Isolated DNA can be used for in vitr...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1993-02-01 00:00:00
abstract：:This report describes a new method for labeling PCR-generated short length (60-120 bp) double-stranded DNA fragments for use as hybridization probes. The method utilizes gene-specific primers identical to those for PCR generation of non-radioactive DNA fragments. Radioactive probes are synthesized by Taq DNA polymeras...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1992-03-01 00:00:00
abstract：:A nonimmune phagemid recombinant antibody fragment (rFab) library was generated with a nominal diversity of 1.16 x 10(7) using the QuikChange Multi Site-Directed Mutagenesis kit. Two degenerate primers spanning the third complementarity-determining region (CDR) loops of the antibody fragment light and heavy chain were...
journal_title：BioTechniques
pub_type：
doi：10.2144/03354st05
更新日期：2003-10-01 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：: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
