Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1997-02-28
1999-07-27
Jones, W. Gary
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
536 231, 536 243, 935 76, 935 77, 935 78, C12Q 168, C07H 2102, C07H 2104, C12N 1500
Patent
active
059288671
ABSTRACT:
Method of isolating exonic gene segments of a eukaryotic gene by preparing a loaded exon trapping vector, bringing the loaded exon trapping vector in an environment in which transcription of DNA to RNA and RNA processing can occur, isolating processed RNA which includes exonic gene segments of the eukaryotic gene and, optionally, preparing a DNA copy of the isolated processed RNA. The loaded exon trapping vector comprises a large-insert DNA vector containing a segmented donor gene construct and, inserted in or close to said segmented donor gene construct, a target DNA fragment of at least 10 kb of eukaryotic genomic DNA. Use of vectors containing large genomic inserts allows to isolate exonic gene segments present in the correct transcriptional orientation as a complete set, without need for reordering individually isolated exons and verifying their continuity from isolated cDNAs. The number of false positives is much reduced, negatives (empty products) can easily be removed, and the system allows isolation of all types of exonic gene fragments, i.e. 5'-first internal and 3'-terminal exons.
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Datson Nicole Anne
Dauwerse Johanes Gregorius
den Dunnen Johan Theodorus
van Ommen Garrit Jan Boudewijn
Jones W. Gary
Rijksuniversiteit Leiden
Whisenant Ethan
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