Magnetic bead purification of M13 DNA sequencing templates

Alderton, Robert P.; Eccleston, Lynne M.; Howe, Roland; Read, Christopher A.; Reeve, Michael A.; Beck, Stephan

doi:10.1016/0003-2697(92)90190-i

Cited by 19 publications

(9 citation statements)

References 6 publications

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“…Random fragments of virion DNA of approximately 2 kb were generated by sonication, end repaired with Klenow fragment of DNA polymerase to produce blunt ends, and used to construct an M13 library (8). Single-stranded sequencing templates were prepared using Escherichia coli TG1, by either previously described techniques (8) or a modified method in microtiter trays (130) employing sodium iodide as the chaotropic agent instead of sodium dodecyl sulfate for cell lysis (3). Doublestranded DNA templates were generated by PCR from the M13 single-stranded template with M13 forward and reverse PCR primers.…”

Section: Methodsmentioning

confidence: 99%

Analysis of the complete DNA sequence of murine cytomegalovirus

Rawlinson

Farrell

Barrell

1996

J Virol

532

275

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The complete DNA sequence of the Smith strain of murine cytomegalovirus (MCMV) was determined from virion DNA by using a whole-genome shotgun approach. The genome has an overall G؉C content of 58.7%, consists of 230,278 bp, and is arranged as a single unique sequence with short (31-bp) terminal direct repeats and several short internal repeats. Significant similarity to the genome of the sequenced human cytomegalovirus (HCMV) strain AD169 is evident, particularly for 78 open reading frames encoded by the central part of the genome. There is a very similar distribution of G؉C content across the two genomes. Sequences toward the ends of the MCMV genome encode tandem arrays of homologous glycoproteins (gps) arranged as two gene families. The left end encodes 15 gps that represent one family, and the right end encodes a different family of 11 gps. A homolog (m144) of cellular major histocompatibility complex (MHC) class I genes is located at the end of the genome opposite the HCMV MHC class I homolog (UL18). G protein-coupled receptor (GCR) homologs (M33 and M78) occur in positions congruent with two (UL33 and UL78) of the four putative HCMV GCR homologs. Counterparts of all of the known enzyme homologs in HCMV are present in the MCMV genome, including the phosphotransferase gene (M97), whose product phosphorylates ganciclovir in HCMVinfected cells, and the assembly protein (M80).

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Section: Methodsmentioning

confidence: 99%

Analysis of the complete DNA sequence of murine cytomegalovirus

Rawlinson

Farrell

Barrell

1996

J Virol

532

275

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“…The process proposed uses a three-phase emulsion containing soluble polymer particles, a monomer phase and water. Nucleic acid separation using magnetic beads is described in (Alderton et al 1992) and in WO/1991/012079 as well as in US 5,523,231 (Amersham). These magnetic beads are able to absorb the nucleic acid after a salt-ethanol precipitation.…”

Section: Commercially Available Magnetic Particlesmentioning

confidence: 99%

“…Another drawback is the danger of mutual contamination of different biological samples, especially if directly neighbouring supports are emptied by the vacuum. However, the last decade shows that DNA purification using magnetic bead technology is suitable for automation systems, and several automated instruments for handling magnetic beads have been developed (Alderton et al 1992;Wahlberg et al 1992;Rolfs and Weber 1994;Fangan et al 1999;Obata et al 2001;Akutsu et al 2004;Vuosku et al 2004).…”

Section: Magnetic Separatorsmentioning

confidence: 99%

Magnetic particles for the separation and purification of nucleic acids

Berensmeier¹

2006

Appl Microbiol Biotechnol

475

296

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Nucleic acid separation is an increasingly important tool for molecular biology. Before modern technologies could be used, nucleic acid separation had been a time- and work-consuming process based on several extraction and centrifugation steps, often limited by small yields and low purities of the separation products, and not suited for automation and up-scaling. During the last few years, specifically functionalised magnetic particles were developed. Together with an appropriate buffer system, they allow for the quick and efficient purification directly after their extraction from crude cell extracts. Centrifugation steps were avoided. In addition, the new approach provided for an easy automation of the entire process and the isolation of nucleic acids from larger sample volumes. This review describes traditional methods and methods based on magnetic particles for nucleic acid purification. The synthesis of a variety of magnetic particles is presented in more detail. Various suppliers of magnetic particles for nucleic acid separation as well as suppliers offering particle-based kits for a variety of different sample materials are listed. Furthermore, commercially available manual magnetic separators and automated systems for magnetic particle handling and liquid handling are mentioned.

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“…DNA extraction using magnetic beads is thought to be suitable for automation systems, and several automated instruments for handling magnetic beads have been developed (Alderton et al, 1992;Fangan et al, 1999;Rolfs and Weber, 1994;Wahlberg et al, 1992). However, they have not been widely used because they cannot achieve a high recovery of magnetic beads.…”

Section: Introductionmentioning

confidence: 99%

Development of an integrated automation system with a magnetic bead‐mediated nucleic acid purification device for genetic analysis and gene manipulation

Akutsu

Tojo

Segawa

et al. 2004

Biotech & Bioengineering

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We have developed an integrated automation system for genetic analysis and gene manipulation. The system, SX-8G Plus, is equipped with an 8-nozzle dispensing unit, a thermal cycler, a cooled reagent reservoir, four tip storage racks, four microplate platforms, buffer reservoirs, an agarose gel electrophoresis unit, a power supply, a pump for exchanging electrophoresis buffer, and a CCD camera. Automation of nucleic acid extraction and purification, the most difficult step in automating genetic analysis and gene manipulation, was realized using magnetic beads with Magtration Technology, which we have previously developed for automating the handling of paramagnetic beads. Using this system, we could perform the automated separation and purification of DNA fragments by agarose gel electrophoresis starting from sample loading. The system would enable the automation of almost all procedures in genetic analysis and gene manipulation.

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Magnetic bead purification of M13 DNA sequencing templates

Cited by 19 publications

References 6 publications

Analysis of the complete DNA sequence of murine cytomegalovirus

Analysis of the complete DNA sequence of murine cytomegalovirus

Magnetic particles for the separation and purification of nucleic acids

Development of an integrated automation system with a magnetic bead‐mediated nucleic acid purification device for genetic analysis and gene manipulation

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