MersV1, Ncbi, Merge, bibRecord, 000235

Sequencing by hybridization: towards an automated sequencing of one million M13 clones arrayed on membranes.

Identifieur interne : 000235 ( Ncbi/Merge ); précédent : 000234; suivant : 000236

Sequencing by hybridization: towards an automated sequencing of one million M13 clones arrayed on membranes.

Auteurs : R. Drmanac [États-Unis] ; S. Drmanac ; I. Labat ; R. Crkvenjakov ; A. Vicentic ; A. Gemmell

Source :

Electrophoresis [ 0173-0835 ] ; 1992.

RBID : pubmed:1451694

Descripteurs français

English descriptors

KwdEn :
- Bacteriophage M13 (genetics), Base Sequence, Cloning, Molecular, DNA (chemistry), Membranes, Artificial, Molecular Sequence Data, Nucleic Acid Hybridization, Oligonucleotide Probes, Phosphorus Radioisotopes, Polymerase Chain Reaction, Robotics.
MESH :
- chemical , chemistry : DNA.
- genetics : Bacteriophage M13.
- Base Sequence, Cloning, Molecular, Membranes, Artificial, Molecular Sequence Data, Nucleic Acid Hybridization, Oligonucleotide Probes, Phosphorus Radioisotopes, Polymerase Chain Reaction, Robotics.

Abstract

An immediately applicable variant of the sequencing by hybridization (SBH) method is under development with the capacity to determine up to 100 million base pairs per year. The proposed method comprises six steps: (i) arraying genomic or cDNA M13 clones in 864-well plates (wells of 2 mm); (ii) preparation of DNA samples for spotting by growth of the M13 clones or by polymerase chain reaction (PCR) of the inserts using standard 96-well plates, or plates having as many as 864 correspondingly smaller wells; (iii) robotic spotting of 13,824 samples on an 8 x 12 cm nylon membrane, or correspondingly more, on up to 6 times larger filters, by offset printing with a 96 or 864 0.4 mm pin device; (iv) hybridization of dotted samples with 200-2000 32P-labeled probes comprising 16-256 10-mers having a common 8-mer, 7-mer, or 6-mer in the middle (20 probes per day, each hybridized with 250,000 dots); (v) scoring hybridization signals of 5 million sample-probe pairs per day using storage phosphor plates; and (vi) computing clone order and partial-to-complete DNA sequences using various heuristic algorithms. Genome sequencing based on a combination of this method and gel sequencing techniques may be significantly more economical than gel methods alone.

DOI: 10.1002/elps.11501301115
PubMed: 1451694

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Le document en format XML

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<term>Base Sequence</term>
<term>Cloning, Molecular</term>
<term>DNA (chemistry)</term>
<term>Membranes, Artificial</term>
<term>Molecular Sequence Data</term>
<term>Nucleic Acid Hybridization</term>
<term>Oligonucleotide Probes</term>
<term>Phosphorus Radioisotopes</term>
<term>Polymerase Chain Reaction</term>
<term>Robotics</term>
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<term>Bactériophage M13 (génétique)</term>
<term>Clonage moléculaire</term>
<term>Données de séquences moléculaires</term>
<term>Hybridation d'acides nucléiques</term>
<term>Membrane artificielle</term>
<term>Radio-isotopes du phosphore</term>
<term>Robotique</term>
<term>Réaction de polymérisation en chaîne</term>
<term>Sondes oligonucléotidiques</term>
<term>Séquence nucléotidique</term>
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<keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>DNA</term>
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<term>Oligonucleotide Probes</term>
<term>Phosphorus Radioisotopes</term>
<term>Polymerase Chain Reaction</term>
<term>Robotics</term>
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<term>Données de séquences moléculaires</term>
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<term>Membrane artificielle</term>
<term>Radio-isotopes du phosphore</term>
<term>Robotique</term>
<term>Réaction de polymérisation en chaîne</term>
<term>Sondes oligonucléotidiques</term>
<term>Séquence nucléotidique</term>
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<front><div type="abstract" xml:lang="en">An immediately applicable variant of the sequencing by hybridization (SBH) method is under development with the capacity to determine up to 100 million base pairs per year. The proposed method comprises six steps: (i) arraying genomic or cDNA M13 clones in 864-well plates (wells of 2 mm); (ii) preparation of DNA samples for spotting by growth of the M13 clones or by polymerase chain reaction (PCR) of the inserts using standard 96-well plates, or plates having as many as 864 correspondingly smaller wells; (iii) robotic spotting of 13,824 samples on an 8 x 12 cm nylon membrane, or correspondingly more, on up to 6 times larger filters, by offset printing with a 96 or 864 0.4 mm pin device; (iv) hybridization of dotted samples with 200-2000 32P-labeled probes comprising 16-256 10-mers having a common 8-mer, 7-mer, or 6-mer in the middle (20 probes per day, each hybridized with 250,000 dots); (v) scoring hybridization signals of 5 million sample-probe pairs per day using storage phosphor plates; and (vi) computing clone order and partial-to-complete DNA sequences using various heuristic algorithms. Genome sequencing based on a combination of this method and gel sequencing techniques may be significantly more economical than gel methods alone.</div>
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Sequencing by hybridization: towards an automated sequencing of one million M13 clones arrayed on membranes.

Sequencing by hybridization: towards an automated sequencing of one million M13 clones arrayed on membranes.

Source :

Descripteurs français

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Links to Exploration step

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

Pour mettre un lien sur cette page dans le réseau Wicri

Pour générer des pages wiki