Specific-primer-directed DNA sequencing using automated fluorescence detection.

Abstract:

:Automated fluorescence-based DNA sequence analysis offers the possibility to undertake very large scale sequencing projects. Directed strategies, such as the specific-primer-directed sequencing approach ('gene walking'), should prove useful in such projects. Described herein is a study involving the use of this approach in conjunction with automated fluorescence detection on a commercial instrument (ABI 370A DNA sequencer). This includes procedures for the rapid chemical synthesis and purification of labeled primers, the design of primer sequences that are compatible with the commercial analysis software, and automated DNA sequence analysis using such primers. A set of four fluorophore-labeled primers can be reliably synthesized in a twenty-four hour period, and greater than 300 nucleotides of analyzed new sequence obtained using this set in an additional twenty-four hours. Scale-up of these procedures to take advantage of the full capabilities of the sequencer is, at present, too slow and costly to be suitable for routine sequencing, and therefore the use of specific-primers is best suited to the closure of gaps in extended sequence produced using random cloning and sequencing strategies.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Kaiser RJ,MacKellar SL,Vinayak RS,Sanders JZ,Saavedra RA,Hood LE

doi

10.1093/nar/17.15.6087

subject

Has Abstract

pub_date

1989-08-11 00:00:00

pages

6087-102

issue

15

eissn

0305-1048

issn

1362-4962

journal_volume

17

pub_type

杂志文章
  • Cloning whole bacterial genomes in yeast.

    abstract::Most microbes have not been cultured, and many of those that are cultivatable are difficult, dangerous or expensive to propagate or are genetically intractable. Routine cloning of large genome fractions or whole genomes from these organisms would significantly enhance their discovery and genetic and functional charact...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkq119

    authors: Benders GA,Noskov VN,Denisova EA,Lartigue C,Gibson DG,Assad-Garcia N,Chuang RY,Carrera W,Moodie M,Algire MA,Phan Q,Alperovich N,Vashee S,Merryman C,Venter JC,Smith HO,Glass JI,Hutchison CA 3rd

    更新日期:2010-05-01 00:00:00

  • Conserved elements with potential to form polymorphic G-quadruplex structures in the first intron of human genes.

    abstract::To understand how potential for G-quadruplex formation might influence regulation of gene expression, we examined the 2 kb spanning the transcription start sites (TSS) of the 18 217 human RefSeq genes, distinguishing contributions of template and nontemplate strands. Regions both upstream and downstream of the TSS are...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkm1138

    authors: Eddy J,Maizels N

    更新日期:2008-03-01 00:00:00

  • Two rotameric forms of open ring 7-methylguanine are present in alkylated polynucleotides.

    abstract::High performance liquid chromatography analysis of imidazole open ring 7-methylguanine, 2-6 diamino-4-hydroxy-5N-methyl-formamidopyrimidine (rom7G), showed two well-separated peaks (fI and fII) of the same magnitude. Rechromatography of each isolated component indicated that they are slowly interconverted to give a 1:...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/12.13.5429

    authors: Boiteux S,Belleney J,Roques BP,Laval J

    更新日期:1984-07-11 00:00:00

  • RaftProt: mammalian lipid raft proteome database.

    abstract::RaftProt (http://lipid-raft-database.di.uq.edu.au/) is a database of mammalian lipid raft-associated proteins as reported in high-throughput mass spectrometry studies. Lipid rafts are specialized membrane microdomains enriched in cholesterol and sphingolipids thought to act as dynamic signalling and sorting platforms....

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gku1131

    authors: Shah A,Chen D,Boda AR,Foster LJ,Davis MJ,Hill MM

    更新日期:2015-01-01 00:00:00

  • BioModels-15 years of sharing computational models in life science.

    abstract::Computational modelling has become increasingly common in life science research. To provide a platform to support universal sharing, easy accessibility and model reproducibility, BioModels (https://www.ebi.ac.uk/biomodels/), a repository for mathematical models, was established in 2005. The current BioModels platform ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkz1055

    authors: Malik-Sheriff RS,Glont M,Nguyen TVN,Tiwari K,Roberts MG,Xavier A,Vu MT,Men J,Maire M,Kananathan S,Fairbanks EL,Meyer JP,Arankalle C,Varusai TM,Knight-Schrijver V,Li L,Dueñas-Roca C,Dass G,Keating SM,Park YM,Buso N

    更新日期:2020-01-08 00:00:00

  • ZAR1 and ZAR2 are required for oocyte meiotic maturation by regulating the maternal transcriptome and mRNA translational activation.

    abstract::Zar1 was one of the earliest mammalian maternal-effect genes to be identified. Embryos derived from Zar1-null female mice are blocked before zygotic genome activation; however, the underlying mechanism remains unclear. By knocking out Zar1 and its homolog Zar2 in mice, we revealed a novel function of these genes in oo...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkz863

    authors: Rong Y,Ji SY,Zhu YZ,Wu YW,Shen L,Fan HY

    更新日期:2019-12-02 00:00:00

  • Dynamics of genetic variation in transcription factors and its implications for the evolution of regulatory networks in Bacteria.

    abstract::The evolution of regulatory networks in Bacteria has largely been explained at macroevolutionary scales through lateral gene transfer and gene duplication. Transcription factors (TF) have been found to be less conserved across species than their target genes (TG). This would be expected if TFs accumulate mutations fas...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkaa162

    authors: Ali F,Seshasayee ASN

    更新日期:2020-05-07 00:00:00

  • Repression of class I transcription by cadmium is mediated by the protein phosphatase 2A.

    abstract::Toxic metals are part of our environment, and undue exposure to them leads to a variety of pathologies. In response, most organisms adapt their metabolism and have evolved systems to limit this toxicity and to acquire tolerance. Ribosome biosynthesis being central for protein synthesis, we analyzed in yeast the effect...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkt335

    authors: Zhou L,Le Roux G,Ducrot C,Chédin S,Labarre J,Riva M,Carles C

    更新日期:2013-07-01 00:00:00

  • Nuclear dynamics of topoisomerase IIβ reflects its catalytic activity that is regulated by binding of RNA to the C-terminal domain.

    abstract::DNA topoisomerase II (topo II) changes DNA topology by cleavage/re-ligation cycle(s) and thus contributes to various nuclear DNA transactions. It is largely unknown how the enzyme is controlled in a nuclear context. Several studies have suggested that its C-terminal domain (CTD), which is dispensable for basal relaxat...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gku640

    authors: Onoda A,Hosoya O,Sano K,Kiyama K,Kimura H,Kawano S,Furuta R,Miyaji M,Tsutsui K,Tsutsui KM

    更新日期:2014-08-01 00:00:00

  • Evolutionary Conserved Motif Finder (ECMFinder) for genome-wide identification of clustered YY1- and CTCF-binding sites.

    abstract::We have developed a new bioinformatics approach called ECMFinder (Evolutionary Conserved Motif Finder). This program searches for a given DNA motif within the entire genome of one species and uses the gene association information of a potential transcription factor-binding site (TFBS) to screen the homologous regions ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkp077

    authors: Kang K,Chung JH,Kim J

    更新日期:2009-04-01 00:00:00

  • Two zinc finger proteins from Xenopus laevis bind the same region of 5S RNA but with different nuclease protection patterns.

    abstract::Immature oocytes from Xenopus laevis contain a 42S ribonucleoprotein particle (RNP) containing 5S RNA, tRNA, a 43 kDa protein, and a 48 kDa protein. A particle containing 5S RNA and the 43 kDa protein (p43-5S) liberated from the 42S particle upon brief treatment with urea can be purified by anion exchange chromatograp...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/19.8.1797

    authors: Sands MS,Bogenhagen DF

    更新日期:1991-04-25 00:00:00

  • DNA binding modes influence Rap1 activity in the regulation of telomere length and MRX functions at DNA ends.

    abstract::The cellular response to DNA double-strand breaks (DSBs) is initiated by the Mre11-Rad50-Xrs2 (MRX) complex that has structural and catalytic functions. MRX association at DSBs is counteracted by Rif2, which is known to interact with Rap1 that binds telomeric DNA through two tandem Myb-like domains. Whether and how Ra...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkz1203

    authors: Bonetti D,Rinaldi C,Vertemara J,Notaro M,Pizzul P,Tisi R,Zampella G,Longhese MP

    更新日期:2020-03-18 00:00:00

  • Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements.

    abstract::Based on parameters governing promoter activity and using regulatory elements of the lac, ara and tet operon transcription control sequences were composed which permit the regulation in Escherichia coli of several gene activities independently and quantitatively. The novel promoter PLtetO-1 allows the regulation of ge...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/25.6.1203

    authors: Lutz R,Bujard H

    更新日期:1997-03-15 00:00:00

  • A library-based method to rapidly analyse chromatin accessibility at multiple genomic regions.

    abstract::Traditional chromatin analysis methods only test one locus at the time or use different templates for each locus, making a standardized analysis of large genomic regions or many co-regulated genes at different loci a difficult task. On the other hand, genome-wide high-resolution mapping of chromatin accessibility empl...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkp037

    authors: Basheer A,Berger H,Reyes-Dominguez Y,Gorfer M,Strauss J

    更新日期:2009-04-01 00:00:00

  • A single zinc finger optimizes the DNA interactions of the nucleocapsid protein of the yeast retrotransposon Ty3.

    abstract::Reverse transcription in retroviruses and retrotransposons requires nucleic acid chaperones, which drive the rearrangement of nucleic acid conformation. The nucleic acid chaperone properties of the human immunodeficiency virus type-1 (HIV-1) nucleocapsid (NC) protein have been extensively studied, and nucleic acid agg...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkr726

    authors: Chaurasiya KR,Geertsema H,Cristofari G,Darlix JL,Williams MC

    更新日期:2012-01-01 00:00:00

  • Two classes of EF1-family translational GTPases encoded by giant viruses.

    abstract::Giant viruses have extraordinarily large dsDNA genomes, and exceptionally, they encode various components of the translation apparatus, including tRNAs, aminoacyl-tRNA synthetases and translation factors. Here, we focused on the elongation factor 1 (EF1) family of viral translational GTPases (trGTPases), using computa...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkz296

    authors: Zinoviev A,Kuroha K,Pestova TV,Hellen CUT

    更新日期:2019-06-20 00:00:00

  • Nucleosomes undergo slow spontaneous gaping.

    abstract::In eukaryotes, DNA is packaged into a basic unit, the nucleosome which consists of 147 bp of DNA wrapped around a histone octamer composed of two copies each of the histones H2A, H2B, H3 and H4. Nucleosome structures are diverse not only by histone variants, histone modifications, histone composition but also through ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkv276

    authors: Ngo TT,Ha T

    更新日期:2015-04-30 00:00:00

  • DNA nicking favors PCR recombination.

    abstract::We attempted to use the polymerase chain reaction (PCR) to monitor in vitro recombination in a plasmid containing directly repeated sequences. Some of the plasmid preparations which had not been exposed to recombination conditions were however found to behave in the PCR test as if they had undergone homologous recombi...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/19.9.2423

    authors: Marton A,Delbecchi L,Bourgaux P

    更新日期:1991-05-11 00:00:00

  • Replication enhancer elements within the open reading frame of tick-borne encephalitis virus and their evolution within the Flavivirus genus.

    abstract::We provide experimental evidence of a replication enhancer element (REE) within the capsid gene of tick-borne encephalitis virus (TBEV, genus Flavivirus). Thermodynamic and phylogenetic analyses predicted that the REE folds as a long stable stem-loop (designated SL6), conserved among all tick-borne flaviviruses (TBFV)...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkr237

    authors: Tuplin A,Evans DJ,Buckley A,Jones IM,Gould EA,Gritsun TS

    更新日期:2011-09-01 00:00:00

  • Organization of the 3'-boundary of the chicken alpha globin gene domain and characterization of a CR 1-specific protein binding site.

    abstract::The sequence of a DNA fragment about 1 Kbp long located at the 3' boundary of the chicken alpha globin gene domain, including the 3'-side matrix attachment point and the site of transcription termination, was determined. It contains a repetitive DNA element and the AT-rich (easily denaturable) DNA segment conserved at...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/18.3.401

    authors: Farache G,Razin SV,Targa FR,Scherrer K

    更新日期:1990-02-11 00:00:00

  • Human fetal globin DNA sequences suggest novel conversion event.

    abstract::DNA sequencing studies of two recently cloned human A gamma globin alleles has revealed a number of base differences which are clustered in the large intron (IVS-2). One allele has a previously undescribed IVS-2 sequence. Most of the allelic differences can be explained as resulting from a gene conversion event involv...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/12.11.4469

    authors: Stoeckert CJ Jr,Collins FS,Weissman SM

    更新日期:1984-06-11 00:00:00

  • Predicted structure and phyletic distribution of the RNA-binding protein Hfq.

    abstract::Hfq, a bacterial RNA-binding protein, was recently shown to contain the Sm1 motif, a characteristic of Sm and LSm proteins that function in RNA processing events in archaea and eukaryotes. In this report, comparative structural modeling was used to predict a three-dimensional structure of the Hfq core sequence. The pr...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkf508

    authors: Sun X,Zhulin I,Wartell RM

    更新日期:2002-09-01 00:00:00

  • See me, feel me: methods to concurrently visualize and manipulate single DNA molecules and associated proteins.

    abstract::Direct visualization of DNA and proteins allows researchers to investigate DNA-protein interactions with great detail. Much progress has been made in this area as a result of increasingly sensitive single-molecule fluorescence techniques. At the same time, methods that control the conformation of DNA molecules have be...

    journal_title:Nucleic acids research

    pub_type: 杂志文章,评审

    doi:10.1093/nar/gkn412

    authors: van Mameren J,Peterman EJ,Wuite GJ

    更新日期:2008-08-01 00:00:00

  • MEPD: a Medaka gene expression pattern database.

    abstract::The Medaka Expression Pattern Database (MEPD) stores and integrates information of gene expression during embryonic development of the small freshwater fish Medaka (Oryzias latipes). Expression patterns of genes identified by ESTs are documented by images and by descriptions through parameters such as staining intensi...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkg017

    authors: Henrich T,Ramialison M,Quiring R,Wittbrodt B,Furutani-Seiki M,Wittbrodt J,Kondoh H,Medaka Expression Pattern Database.

    更新日期:2003-01-01 00:00:00

  • G-quadruplex recognition and remodeling by the FANCJ helicase.

    abstract::Guanine rich nucleic acid sequences can form G-quadruplex (G4) structures that interfere with DNA replication, repair and RNA transcription. The human FANCJ helicase contributes to maintaining genomic integrity by promoting DNA replication through G4-forming DNA regions. Here, we combined single-molecule and ensemble ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkw574

    authors: Wu CG,Spies M

    更新日期:2016-10-14 00:00:00

  • A screen of chemical modifications identifies position-specific modification by UNA to most potently reduce siRNA off-target effects.

    abstract::Small interfering RNAs (siRNAs) are now established as the preferred tool to inhibit gene function in mammalian cells yet trigger unintended gene silencing due to their inherent miRNA-like behavior. Such off-target effects are primarily mediated by the sequence-specific interaction between the siRNA seed regions (posi...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkq341

    authors: Bramsen JB,Pakula MM,Hansen TB,Bus C,Langkjær N,Odadzic D,Smicius R,Wengel SL,Chattopadhyaya J,Engels JW,Herdewijn P,Wengel J,Kjems J

    更新日期:2010-09-01 00:00:00

  • Ultraviolet light irradiation of PM2 superhelical DNA.

    abstract::Superhelical PM2 DNA can be photochemically modified by u.v. irradiation. The variation of S20,w with dose shows the following characteristics. There is a linear increase from 28 to 31s produced by a low dose of u.v. irradiation (4,000 ergs/mm2). A plateau in S20,w occurs between 4,000 and 10,000 ergs/mm2. The S20,w t...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/4.5.1243

    authors: Woodworth-Gutai M,Lebowitz J,Kato AC,Denhardt DT

    更新日期:1977-01-01 00:00:00

  • The Aspergillus Genome Database, a curated comparative genomics resource for gene, protein and sequence information for the Aspergillus research community.

    abstract::The Aspergillus Genome Database (AspGD) is an online genomics resource for researchers studying the genetics and molecular biology of the Aspergilli. AspGD combines high-quality manual curation of the experimental scientific literature examining the genetics and molecular biology of Aspergilli, cutting-edge comparativ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkp751

    authors: Arnaud MB,Chibucos MC,Costanzo MC,Crabtree J,Inglis DO,Lotia A,Orvis J,Shah P,Skrzypek MS,Binkley G,Miyasato SR,Wortman JR,Sherlock G

    更新日期:2010-01-01 00:00:00

  • Systematic use of synthetic 5'-UTR RNA structures to tune protein translation improves yield and quality of complex proteins in mammalian cell factories.

    abstract::Predictably regulating protein expression levels to improve recombinant protein production has become an important tool, but is still rarely applied to engineer mammalian cells. We therefore sought to set-up an easy-to-implement toolbox to facilitate fast and reliable regulation of protein expression in mammalian cell...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkaa847

    authors: Eisenhut P,Mebrahtu A,Moradi Barzadd M,Thalén N,Klanert G,Weinguny M,Sandegren A,Su C,Hatton D,Borth N,Rockberg J

    更新日期:2020-11-18 00:00:00

  • The scaRNA2 is produced by an independent transcription unit and its processing is directed by the encoding region.

    abstract::The C/D box scaRNA2 is predicted to guide specific 2'-O-methylation of U2 snRNA. In contrast to other SCARNA genes, SCARNA2 appears to be independently transcribed. By transient expression of SCARNA2-reporter gene constructs, we have demonstrated that this gene is transcribed by RNA polymerase II and that the promoter...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkp988

    authors: Gérard MA,Myslinski E,Chylak N,Baudrey S,Krol A,Carbon P

    更新日期:2010-01-01 00:00:00