Glass bead purification of plasmid template DNA for high throughput sequencing of mammalian genomes

Nucleic Acids Research
Debra A DederichMichael L Metzker

Abstract

To meet the new challenge of generating the draft sequences of mammalian genomes, we describe the development of a novel high throughput 96-well method for the purification of plasmid DNA template using size-fractionated, acid-washed glass beads. Unlike most previously described approaches, the current method has been designed and optimized to facilitate the direct binding of alcohol-precipitated plasmid DNA to glass beads from alkaline lysed bacterial cells containing the insoluble cellular aggregate material. Eliminating the tedious step of separating the cleared lysate significantly simplifies the method and improves throughput and reliability. During a 4 month period of 96-capillary DNA sequencing of the Rattus norvegicus genome at the Baylor College of Medicine Human Genome Sequencing Center, the average success rate and read length derived from >1 800 000 plasmid DNA templates prepared by the direct lysis/glass bead method were 82.2% and 516 bases, respectively. The cost of this direct lysis/glass bead method in September 2001 was approximately 10 cents per clone, which is a significant cost saving in high throughput genomic sequencing efforts.

References

Feb 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·B Vogelstein, D Gillespie
Nov 24, 1979·Nucleic Acids Research·H C Birnboim, J Doly
Jan 15, 1980·Analytical Biochemistry·C W Chen, C A Thomas
Jun 1, 1981·Analytical Biochemistry·D S Holmes, M Quigley
Oct 25, 1994·Nucleic Acids Research·T L HawkinsC Santillan
Apr 5, 1996·Analytical Biochemistry·B AnderssonR A Gibbs
Dec 15, 1996·Nucleic Acids Research·W L NgL Hood
Nov 26, 1999·Nucleic Acids Research·M A MarraR H Waterston
Mar 14, 2001·Science·E Marshall
Jul 4, 2001·Genome Research·C J ElkinT L Hawkins

❮ Previous
Next ❯

Citations

Dec 17, 2009·Journal of Biomedicine & Biotechnology·Siun Chee Tan, Beow Chin Yiap
Dec 3, 2013·Analytical Biochemistry·Ji Hyun MinYoung Keun Kim
Jun 15, 2007·Molecular and Biochemical Parasitology·Taisei KikuchiJohn T Jones
Dec 20, 2005·Molecular Phylogenetics and Evolution·W Brian SimisonJ L Boore
May 21, 2005·Molecular Phylogenetics and Evolution·J Robert MaceyJeffrey L Boore
Oct 14, 2014·Expert Review of Vaccines·Alex Xenopoulos, Priyabrata Pattnaik
May 17, 2017·Chemistry, an Asian Journal·Xuan-Hung PhamBong-Hyun Jun
Mar 15, 2019·BioMed Research International·Aisha Nawaf Al BalawiHamad A Al-Lohedan
Jun 24, 2021·Scientific Reports·Fereshte DamavandiGane Ka-Shu Wong

❮ Previous
Next ❯

Related Concepts

Related Feeds

Brain Lower Grade Glioma

Low grade gliomas in the brain form from oligodendrocytes and astrocytes and are the slowest-growing glioma in adults. Discover the latest research on these brain tumors here.