Abstract:
:The need for expeditious and inexpensive methods for high-throughput DNA sequencing has been highlighted by the accelerated pace of genome DNA sequencing over the past year. At the Joint Genome Institute, the throughput in terms of high-quality bases per day has increased over 20-fold during the past 18 mo, reaching an average of 18.3 million bases per day. To support this unprecedented scaleup, we developed an inexpensive automated method for the isolation and purification of double-stranded plasmid DNA clones for sequencing that is tailored to meet the more stringent needs of the newer capillary electrophoresis DNA sequencing machines. The protocol is based on the magnetic bead method of solid phase reversible immobilization that has been automated by using a CRS-based robotic system. The method described here has enabled us to meet our increases in production while reducing labor and materials costs significantly.
journal_name
Genome Resjournal_title
Genome researchauthors
Elkin CJ,Richardson PM,Fourcade HM,Hammon NM,Pollard MJ,Predki PF,Glavina T,Hawkins TLdoi
10.1101/gr.167801subject
Has Abstractpub_date
2001-07-01 00:00:00pages
1269-74issue
7eissn
1088-9051issn
1549-5469journal_volume
11pub_type
杂志文章相关文献
GENOME RESEARCH文献大全abstract::Advances in high-throughput technologies, such as ChIP-chip, and the completion of human and mouse genomic sequences now allow analysis of the mechanisms of gene regulation on a systems level. In this study, we have developed a computational genomics approach (termed ChIPModules), which begins with experimentally dete...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.5520206
更新日期:2006-12-01 00:00:00
abstract::We have cloned the human gene encoding the transcription factor T. T protein is vital for the formation of posterior mesoderm and axial development in all vertebrates. Brachyury mutant mice, which lack T protein, die in utero with abnormal notochord, posterior somites, and allantois. We have identified human T genomic...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.6.3.226
更新日期:1996-03-01 00:00:00
abstract::Genomic imprinting is a developmentally important mechanism that involves both differential DNA methylation and allelic histone modifications. Through detailed comparative characterization, a large imprinted domain mapping to chromosome 7q21 in humans and proximal chromosome 6 in mice was redefined. This domain is org...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.077115.108
更新日期:2008-08-01 00:00:00
abstract::Interactions mediated by cell surface receptors initiate important instructive signaling cues but can be difficult to detect in biochemical assays because they are often highly transient and membrane-embedded receptors are difficult to solubilize in their native conformation. Here, we address these biochemical challen...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.231183.117
更新日期:2018-09-01 00:00:00
abstract::Short tandem repeats (STRs) have a wide range of applications, including medical genetics, forensics, and genetic genealogy. High-throughput sequencing (HTS) has the potential to profile hundreds of thousands of STR loci. However, mainstream bioinformatics pipelines are inadequate for the task. These pipelines treat S...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.135780.111
更新日期:2012-06-01 00:00:00
abstract::Spatially distinct gene expression profiles in neural stem cells (NSCs) are a prerequisite to the formation of neuronal diversity, but how these arise from the regulatory interactions between chromatin accessibility and transcription factor activity has remained unclear. Here, we demonstrate that, despite their distin...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.203513.115
更新日期:2016-07-01 00:00:00
abstract::Levels of diversity vary across the human genome. This variation is caused by two forces: differences in mutation rates and the differential impact of natural selection. Pertinent to the question of the relative importance of these two forces is the observation that both diversity within species and interspecies diver...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.3461105
更新日期:2005-09-01 00:00:00
abstract::RNA-seq protocols that focus on transcript termini are well suited for applications in which template quantity is limiting. Here we show that, when applied to end-sequencing data, analytical methods designed for global RNA-seq produce computational artifacts. To remedy this, we created the End Sequence Analysis Toolki...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.207902.116
更新日期:2016-10-01 00:00:00
abstract::Fetal liver intriguingly consists of hepatic parenchymal cells and hematopoietic stem/progenitor cells. Human fetal liver aged 22 wk of gestation (HFL22w) corresponds to the turning point between immigration and emigration of the hematopoietic system. To gain further molecular insight into its developmental and functi...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.175501
更新日期:2001-08-01 00:00:00
abstract::We developed a microarray platform for PCR amplification-independent expression profiling of minute samples. A novel scanning system combined with specialized biochips enables detection down to individual fluorescent oligonucleotide molecules specifically hybridized to their complementary sequence over the entire bioc...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.4999906
更新日期:2006-08-01 00:00:00
abstract::We have developed a novel quantitative method for rapidly assessing the CpG methylation density of a DNA region in mammalian cells. After bisulfite modification of genomic DNA, the region of interest is PCR amplified with primers containing two dam sites (GATC). The purified PCR products are then incubated with 14C-la...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.202501
更新日期:2002-01-01 00:00:00
abstract::Microbial eukaryotes are integral components of natural microbial communities, and their inclusion is critical for many ecosystem studies, yet the majority of published metagenome analyses ignore eukaryotes. In order to include eukaryotes in environmental studies, we propose a method to recover eukaryotic genomes from...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.228429.117
更新日期:2018-04-01 00:00:00
abstract::Mutation rates of microsatellites vary greatly among loci. The causes of this heterogeneity remain largely enigmatic yet are crucial for understanding numerous human neurological diseases and genetic instability in cancer. In this first genome-wide study, the relative contributions of intrinsic features and regional g...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.7113408
更新日期:2008-01-01 00:00:00
abstract::Aberrations of protein-coding genes are a focus of cancer genomics; however, the impact of oncogenes on expression of the ~50% of transcripts without protein-coding potential, including long noncoding RNAs (lncRNAs), has been largely uncharacterized. Activating mutations in the BRAF oncogene are present in >70% of mel...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.140061.112
更新日期:2012-06-01 00:00:00
abstract::A large fraction of the genes from sequenced organisms are of unknown function. This limits biological insight, and for pathogenic microorganisms hampers the development of new approaches to battle infections. There is thus a great need for novel strategies that link genotypes to phenotypes for microorganisms. We desc...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.137430.112
更新日期:2012-12-01 00:00:00
abstract::Variation in the composition of the human oral microbiome in health and disease has been observed. We have characterized inter- and intra-individual variation of microbial communities of 107 individuals in one of the largest cohorts to date (264 saliva samples), using culture-independent 16S rRNA pyrosequencing. We ex...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.140608.112
更新日期:2012-11-01 00:00:00
abstract::Gene expression can be regulated at multiple levels, but it is not known if and how there is broad coordination between regulation at the transcriptional and post-transcriptional levels. Transcription factors and chromatin regulate gene expression transcriptionally, whereas microRNAs (miRNAs) are small regulatory RNAs...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.238311.118
更新日期:2019-02-01 00:00:00
abstract::DNA methylation is found in many eukaryotes, but its function is still controversial. We have studied the methylation of plant and animal genomes using a PCR-based technique amenable for high throughput. Repetitive elements are methylated in both organisms, but whereas most mammalian exons are methylated, plant exons ...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.1784803
更新日期:2003-12-01 00:00:00
abstract::Many CpG islands have tissue-dependent and differentially methylated regions (T-DMRs) in normal cells and tissues. To elucidate how DNA methyltransferases (Dnmts) participate in methylation of the genomic components, we investigated the genome-wide DNA methylation pattern of the T-DMRs with Dnmt1-, Dnmt3a-, and/or Dnm...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.2431504
更新日期:2004-09-01 00:00:00
abstract::Sequencing of the human Y chromosome has uncovered the peculiarities of the genomic organization of a heterogametic sex chromosome of old evolutionary age, and has led to many insights into the evolutionary changes that occurred during its long history. We have studied the genomic organization of the medaka fish Y chr...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.5016106
更新日期:2006-07-01 00:00:00
abstract::While metagenomics has emerged as a technology of choice for analyzing bacterial populations, the assembly of metagenomic data remains challenging, thus stifling biological discoveries. Moreover, recent studies revealed that complex bacterial populations may be composed from dozens of related strains, thus further amp...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.213959.116
更新日期:2017-05-01 00:00:00
abstract::The genomic alterations associated with cancers are numerous and varied, involving both isolated and large-scale complex genomic rearrangements (CGRs). Although the underlying mechanisms are not well understood, CGRs have been implicated in tumorigenesis. Here, we introduce CouGaR, a novel method for characterizing th...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.211201.116
更新日期:2017-01-01 00:00:00
abstract::As the Human Genome Project moves into its sequencing phase, a serious problem has arisen. The same problem has been increasingly vexing in the closing phase of the Caenorhabditis elegans project. The difficulty lies in sequencing efficiently through certain regions in which the templates (DNA substrates for the seque...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.8.5.562
更新日期:1998-05-01 00:00:00
abstract::Integrating the genotype with epigenetic marks holds the promise of better understanding the biology that underlies the complex interactions of inherited and environmental components that define the developmental origins of a range of disorders. The quality of the in utero environment significantly influences health o...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.171439.113
更新日期:2014-07-01 00:00:00
abstract::Eukaryotic translation initiation involves preinitiation ribosomal complex 5'-to-3' directional probing of mRNA for codons suitable for starting protein synthesis. The recognition of codons as starts depends on the codon identity and on its immediate nucleotide context known as Kozak context. When the context is weak ...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.257352.119
更新日期:2020-07-01 00:00:00
abstract::Tissue development and function are exquisitely dependent on proper regulation of gene expression, but it remains controversial whether the genomic signals controlling this process are subject to strong selective constraint. While some studies show that highly constrained noncoding regions act to enhance transcription...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.083089.108
更新日期:2008-12-01 00:00:00
abstract::Transposable elements (TEs) proliferate within the genome of their host, which responds by silencing them epigenetically. Much is known about the mechanisms of silencing in plants, particularly the role of siRNAs in guiding DNA methylation. In contrast, little is known about siRNA targeting patterns along the length o...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.193763.115
更新日期:2016-02-01 00:00:00
abstract::Careful titration of Vent polymerase activity allows efficient amplification of full-length plasmids (12 kb). The high processivity and fidelity of this enzyme made oligonucleotide-directed site-specific mutagenesis of plasmids a straight-forward process. Using only two primers, a mutagenic and a complementary, single...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.5.4.404
更新日期:1995-11-01 00:00:00
abstract::The regulation of gene expression is mediated at the transcriptional level by enhancer regions that are bound by sequence-specific transcription factors (TFs). Recent studies have shown that the in vivo binding sites of single TFs differ between developmental or cellular contexts. How this context-specific binding is ...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.132811.111
更新日期:2012-10-01 00:00:00
abstract::Through comparative studies of the model organism Arabidopsis thaliana and its close relative Brassica oleracea, we have identified conserved regions that represent potentially functional sequences overlooked by previous Arabidopsis genome annotation methods. A total of 454,274 whole genome shotgun sequences covering ...
journal_title:Genome research
pub_type: 杂志文章
doi:10.1101/gr.3176505
更新日期:2005-04-01 00:00:00