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MULTIPLEX DNA SEQUENCING [1 record]
Record 1 - internal organization data 1991-07-16
Record 1, English
Record 1, Subject field(s)
- Genetics
Record 1, Main entry term, English
- multiplex DNA sequencing
1, record 1, English, multiplex%20DNA%20sequencing
correct
Record 1, Abbreviations, English
Record 1, Synonyms, English
Record 1, Textual support, English
Record number: 1, Textual support number: 1 CONT
Multiplex DNA sequencing: The increasing demand for DNA sequences can be met by replacement of each DNA sample in a device with a mixture of N samples so that the normal throughput is increased by a factor of N. Such a method is described. In order to separate the sequence information at the end of the processing, the DNA molecules of interest are ligated to a set of oligonucleotide "tags" at the beginning. The tagged DNA molecules are pooled, amplified, and chemically fragmented in 96-well plates. The resulting reaction products are fractionated by size on sequencing gels and transferred to nylon membranes. These membranes are then probed as many times as there are types of tags in the original pools, producing, in each cycle of probing, autoradiographs similar to those from standard DNA sequencing methods. 1, record 1, English, - multiplex%20DNA%20sequencing
Record 1, French
Record 1, Domaine(s)
- Génétique
Record 1, Main entry term, French
- méthode de séquençage multiplex 1, record 1, French, m%C3%A9thode%20de%20s%C3%A9quen%C3%A7age%20multiplex
Record 1, Abbreviations, French
Record 1, Synonyms, French
Record 1, Textual support, French
Record number: 1, Textual support number: 1 CONT
La méthode de séquençage multiplex : Plusieurs bibliothèques génomiques sont construites dans des vecteurs plasmidiques contenant chacun une séquence nucléotidique particulière, séquence cible spécifique. Des lots de 20 colonies provenant de chacune des 20 bibliothèques sont alors formés et les plasmides de chaque lot sont purifiés. La digestion enzymatique permet d'exciser les inserts contenant les fragments d'ADN génomique flanqués de leurs deux séquences cibles. Les opérations de séquençage sont alors effectuées selon la méthode chimique de Maxam et Gilbert, et les fragments obtenus sont séparés sur gel de polyacrylamide par électrophorèse. Une sonde correspondant à la séquence caractéristique du vecteur est utilisée pour révéler les fragments issus de ce clone. 1, record 1, French, - m%C3%A9thode%20de%20s%C3%A9quen%C3%A7age%20multiplex
Record 1, Spanish
Record 1, Textual support, Spanish
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