Nucleic Acids Research, Vol 24, Issue 20 3879-3886, Copyright © 1996 by Oxford University Press
PJ Oefner, SP Hunicke-Smith, L Chiang, F Dietrich, J Mulligan and RW Davis
Based on a high-performance liquid chromatographic pump, we have built a
device that allows recirculation of DNA through a 63-microm orifice with
ensuing fractionation to a minimum fragment size of approximately 300 base
pairs. Residence time of the DNA fragments in the converging flow created
by a sudden contraction was found to be sufficiently long to allow
extension of the DNA molecules into a highly extended conformation and,
hence, breakage to occur at midpoint. In most instances, 30 passages
sufficed to obtain a narrow size distribution, with >90% of the
fragments lying within a 2-fold size distribution. The shear rate required
to achieve breakage was found to be inversely proportional to the 1.0 power
of the molecular weight. Compared with a restriction digest, up to 40% of
all fragments could be cloned directly, with only marginal improvements in
cloning efficiency having been observed upon prior end repair with Klenow,
T4 polymerase or T4 polynucleotide kinase. Sequencing revealed a fairly
random distribution of the fragments.
ARTICLES
Efficient random subcloning of DNA sheared in a recirculating point- sink flow system
Department of Biochemistry, Stanford University, CA, USA. oefner@genome.stanford.edu
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  J. P. Monteiro, K. V. Clemons, L. F. Mirels, J. A. Coller Jr, T. D. Wu, J. Shankar, C. R. Lopes, and D. A. Stevens Genomic DNA microarray comparison of gene expression patterns in Paracoccidioides brasiliensis mycelia and yeasts in vitro Microbiology, August 1, 2009; 155(8): 2795 - 2808. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. S. Hawkins, H. Kim, J. D. Nason, R. A. Wing, and J. F. Wendel Differential lineage-specific amplification of transposable elements is responsible for genome size variation in Gossypium Genome Res., October 1, 2006; 16(10): 1252 - 1261. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Schneider, U. Dobrindt, H. Bruggemann, G. Nagy, B. Janke, G. Blum-Oehler, C. Buchrieser, G. Gottschalk, L. Emody, and J. Hacker The Pathogenicity Island-Associated K15 Capsule Determinant Exhibits a Novel Genetic Structure and Correlates with Virulence in Uropathogenic Escherichia coli Strain 536 Infect. Immun., October 1, 2004; 72(10): 5993 - 6001. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Grozdanov, C. Raasch, J. Schulze, U. Sonnenborn, G. Gottschalk, J. Hacker, and U. Dobrindt Analysis of the Genome Structure of the Nonpathogenic Probiotic Escherichia coli Strain Nissle 1917 J. Bacteriol., August 15, 2004; 186(16): 5432 - 5441. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Marino-Ramirez, J. L. Minor, N. Reading, and J. C. Hu Identification and Mapping of Self-Assembling Protein Domains Encoded by the Escherichia coli K-12 Genome by Use of {lambda} Repressor Fusions J. Bacteriol., March 1, 2004; 186(5): 1311 - 1319. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. L. Toffaletti, M. Del Poeta, T. H. Rude, F. Dietrich, and J. R. Perfect Regulation of cytochrome c oxidase subunit 1 (COX1) expression in Cryptococcus neoformans by temperature and host environment Microbiology, April 1, 2003; 149(4): 1041 - 1049. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. D. Larbig, A. Christmann, A. Johann, J. Klockgether, T. Hartsch, R. Merkl, L. Wiehlmann, H.-J. Fritz, and B. Tummler Gene Islands Integrated into tRNAGly Genes Confer Genome Diversity on a Pseudomonas aeruginosa Clone J. Bacteriol., December 1, 2002; 184(23): 6665 - 6680. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
U. Dobrindt, G. Blum-Oehler, G. Nagy, G. Schneider, A. Johann, G. Gottschalk, and J. Hacker Genetic Structure and Distribution of Four Pathogenicity Islands (PAI I536 to PAI IV536) of Uropathogenic Escherichia coli Strain 536 Infect. Immun., November 1, 2002; 70(11): 6365 - 6372. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Grozdanov, U. Zahringer, G. Blum-Oehler, L. Brade, A. Henne, Y. A. Knirel, U. Schombel, J. Schulze, U. Sonnenborn, G. Gottschalk, et al. A Single Nucleotide Exchange in the wzy Gene Is Responsible for the Semirough O6 Lipopolysaccharide Phenotype and Serum Sensitivity of Escherichia coli Strain Nissle 1917 J. Bacteriol., November 1, 2002; 184(21): 5912 - 5925. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. B. Lengeler, D. S. Fox, J. A. Fraser, A. Allen, K. Forrester, F. S. Dietrich, and J. Heitman Mating-Type Locus of Cryptococcus neoformans: a Step in the Evolution of Sex Chromosomes Eukaryot. Cell, October 1, 2002; 1(5): 704 - 718. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
U. Dobrindt, G. Blum-Oehler, T. Hartsch, G. Gottschalk, E. Z. Ron, R. Funfstuck, and J. Hacker S-Fimbria-Encoding Determinant sfaI Is Located on Pathogenicity Island III536 of Uropathogenic Escherichia coli Strain 536 Infect. Immun., July 1, 2001; 69(7): 4248 - 4256. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. R. Thorstenson, S. P. Hunicke-Smith, P. J. Oefner, and R. W. Davis An Automated Hydrodynamic Process for Controlled, Unbiased DNA Shearing Genome Res., August 1, 1998; 8(8): 848 - 855. [Abstract] [Full Text]
|
|