Abstract:
:We describe a simple and inexpensive method of performing sequencing reactions for 24 single-strand M13 DNA clones in microtiter plates. To simplify elevated temperature incubations during sequencing reactions, two heating blocks were designed to accommodate microtiter plates and fit within common laboratory heating modules. With only slight modification of standard fluorescent and radioisotopic sequencing methods, the sequencing reactions for 24 clones can be done in as little as 40 minutes.
journal_name
Biotechniquesjournal_title
BioTechniquesauthors
Koop BF,Wilson RK,Chen C,Halloran N,Sciammis R,Hood L,Lindelien JWsubject
Has Abstractpub_date
1990-07-01 00:00:00pages
32, 34-7issue
1eissn
0736-6205issn
1940-9818journal_volume
9pub_type
杂志文章相关文献
BIOTECHNIQUES文献大全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::Radiolabeled proteins are used in a variety of laboratory applications as well as in radioimmunotherapy. This review focuses on methods that utilize genetic engineering to introduce exogenous phosphorylation sites into proteins. Protein kinase substrate sites can be introduced into target proteins to serve as tags for...
journal_title:BioTechniques
pub_type: 杂志文章,评审
doi:
更新日期:2002-10-01 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
pub_type: 杂志文章
doi:10.2144/02326bc03
更新日期:2002-06-01 00:00:00
abstract::Sample automation and management is increasingly important as the number and size of population-scale and high-throughput projects grow. This is particularly the case in large-scale population studies where sample size is far outpacing the commonly used 96-well plate format. To facilitate management and transfer of sa...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/btn-2018-0090
更新日期:2018-12-01 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::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::Loop-mediated isothermal amplification (LAMP), a novel gene amplification method, enables the synthesis of larger amounts of both DNA and a visible byproduct--namely, magnesium pyrophosphate--without thermal cycling. A positive reaction is indicated by the turbidity of the reaction solution or the color change after a...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000113072
更新日期:2009-03-01 00:00:00
abstract::MicroRNAs (miRNAs) are ~22-nucleotide-long small non-coding RNAs that regulate the expression of protein-coding genes by base pairing to partially complementary target sites, preferentially located in the 3´ untranslated region (UTR) of target mRNAs. The expression and function of miRNAs have been extensively studied ...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000114574
更新日期:2017-08-01 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::A head-to-tail trimer of the SV40 Bcl I-Bam H1 DNA fragment, specifying polyadenylation of RNA transcripts, was cloned as a cassette flanked by multiple restriction sites. Insertion of the trimer into several expression vectors efficiently prevented spurious expression of reporter genes resulting from transcriptional ...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1989-03-01 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::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::Here we introduce a modified antibody staining method that uses up to 80% less antibody for flow cytometry. We demonstrate this method for the detection of antigens expressed at high, moderate, or low levels in mouse and rat lymphocytes as well as mouse mammary epithelial cells. We obtained reproducibly accurate resul...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/0000113854
更新日期:2012-07-01 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::The increasing interest in manipulating neural circuits in developing brains has created a demand for reliable and accurate methods for delivering viruses to newborn mice. Here we describe a novel 3D-printed mouse neonatal stereotaxic adaptor for intracerebral viral injection that provides enhanced precision and relia...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/btn-2020-0050
更新日期:2020-10-01 00:00:00
abstract::A method is described for preparing mutants with multiple, site-directed mutations by ordered coupling of PCR-generated fragments catalyzed by a thermostable DNA ligase. Annealing of the sense strands of the fragments to a single-stranded (antisense) template created a full-length sense strand leaving only nicks that ...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1993-07-01 00:00:00
abstract::A simple and rapid procedure for recovering the denaturing effect of methylmercuric hydroxide in agarose gel electrophoresis is described. The procedure consisted of the treatment of the commercial methylmercuric hydroxide solutions with Amberlite, a mixture of anion- and cation-exchange resins. This treatment greatly...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1992-04-01 00:00:00
abstract::Determining the primary sequences of informational macromolecules is no longer a limiting factor for our ability to completely understand the biological functioning of cells and organisms. Similarly, our understanding of transcriptional regulation (transcriptomics) has been greatly enhanced by the availability of micr...
journal_title:BioTechniques
pub_type: 杂志文章,评审
doi:10.2144/000113137
更新日期:2009-04-01 00:00:00
abstract::Autoantibodies directed against the 68-kDa (U1) ribonucleoprotein antigen are mainly found in sera of patients with mixed connective tissue disease. The corresponding cDNA was fragmented into four regions coding for the major antigenic epitopes A', B', C' and D'. All the epitopes were subcloned and expressed as fusion...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1991-09-01 00:00:00
abstract::A procedure for amplification by PCR of reproducible allele markers for amplified fragment length polymorphism (Amp-FLP) analysis is presented. We have prepared markers for the allelic products of the VNTR loci D1S80 (MCT118) and D17S30 (YNZ22) and for the hypervariable VNTR locus close to the 3' end of the apolipopro...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1992-01-01 00:00:00
abstract::A technique, Replacement PCR Mutagenesis, was developed to replace one immunoglobulin variable region (V) in a M13 phage cassette with a different, homologous V. This allows the use of the same mutagenesis and subsequent expression vectors for many V regions or V segments. The method combines PCR of V fragments and in...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1992-01-01 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::We have developed a rapid [3H]colchicine competition-binding scintillation proximity assay (SPA) to evaluate antimitotic compounds that bind to the colchicine-binding site on tubulin. The premise of our assay is that compounds will compete with radiolabeled colchicine for the tubulin-binding domain. Biotin-labeled tub...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/00291rr02
更新日期:2000-07-01 00:00:00
abstract::We report here an improved method for analyzing protein surface expression utilizing a cold-adapted trypsin. Preservation of activity of the enzyme at 0-4°C permits modification of the protease method of surface analysis to temperatures at which trafficking of mammalian plasmalemmal proteins is blocked. This is an imp...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000113651
更新日期:2011-04-01 00:00:00
abstract::The use of affinity-based tools has become invaluable as a platform for basic research and in the development of drugs and diagnostics. Applications include affinity chromatography and affinity tag fusions for efficient purification of proteins as well as methods to probe the protein network interactions on a whole-pr...
journal_title:BioTechniques
pub_type: 杂志文章,评审
doi:10.2144/000112803
更新日期:2008-04-01 00:00:00
abstract::Cycling probe technology (CPT) is a simple signal amplification method for the detection of specific target DNA sequences. CPT uses a chimeric DNA-RNA-DNA probe that is cut by RNase H when bound to its complementary target sequence. In this study, a hybridization assay was developed to detect biotinylated CPT products...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/00286st05
更新日期:2000-06-01 00:00:00
abstract::Few studies have focused on the significance of ras protein levels in human malignancy, in part because of the inherent difficulty in quantitation of the ras gene product. We have developed a method for the enzymatic determination of the ras gene product and have used this method for the quantitation of ras gene produ...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1990-08-01 00:00:00
abstract::Multiplex Manager 1.0 is a user-friendly cross-platform program that designs efficient combinations of existing genetic marker loci into multiplex polymerase chain reactions and optimizes using prior marker information. The program has the flexibility to solve two design problems: combining all markers into the smalle...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000113156
更新日期:2009-06-01 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::Duracryl is a mechanically strong and elastic acrylamide-based matrix, useful for a wide variety of electrophoretic applications. The matrix is stable as a refrigerated solution for one year. Upon addition of appropriate catalysts, Duracryl forms a polymer-reinforced polyacrylamide gel matrix suitable for electrophore...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1992-04-01 00:00:00