Nucleotide sequencing double-stranded plasmids with primers selected from a nonamer library
Nonamer primers, selected from a nonamer library, were tested by sequencing two plasmid subclones containing known insert sequences. These sequences were scanned (nonamer-mapped) against the 2391-member nonamer library to identify all members that share a 100% match at only one site. A total of 59 n...
| Veröffentlicht in: | BioTechniques. - 1988. - 17(1994), 3 vom: 23. Sept., Seite 536-7, 540-4 |
|---|---|
| 1. Verfasser: | |
| Weitere Verfasser: | , , , |
| Format: | Aufsatz |
| Sprache: | English |
| Veröffentlicht: |
1994
|
| Zugriff auf das übergeordnete Werk: | BioTechniques |
| Schlagworte: | Journal Article DNA Primers Globins 9004-22-2 |
| Zusammenfassung: | Nonamer primers, selected from a nonamer library, were tested by sequencing two plasmid subclones containing known insert sequences. These sequences were scanned (nonamer-mapped) against the 2391-member nonamer library to identify all members that share a 100% match at only one site. A total of 59 nonamers were tested using a slightly modified T7 polymerase sequencing procedure for double-stranded DNA. The success rate for nonamer primed reactions was about 60%, and single-stranded coverage was obtained for approximately 90% of each plasmid insert. The results presented demonstrate that a nonamer library, with as few as 2391 members, can greatly aid the completion of many sequencing projects by reducing the number of required custom primers. With the development of a technique for the rapid identification of all useful library primers for a particular sequencing project, one could envision a high-throughput shotgun-type sequencing procedure that would not require large numbers of subclones |
|---|---|
| Beschreibung: | Date Completed 10.02.1995 Date Revised 01.05.2006 published: Print Citation Status MEDLINE |
| ISSN: | 0736-6205 |