SCREENING OF BACTERIAL ARTIFICIAL CHROMOSOME CLONES WITH AFLP-BASED DNA FINGERPRINTING Skrining Klon-Klon BAC Menggunakan DNA-Fingerprinting Berbasis AFLP

Jamsari , Jamsari and Nitzt , Inke and Büttner, Stella Marie Reamon and Jung, Christian (2010) SCREENING OF BACTERIAL ARTIFICIAL CHROMOSOME CLONES WITH AFLP-BASED DNA FINGERPRINTING Skrining Klon-Klon BAC Menggunakan DNA-Fingerprinting Berbasis AFLP. Jurnal Stigma, XII (4). ISSN 0853-3776

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Abstract

With the aim of male-determining gene isolation from asparagus, a Map-based Cloning (MBC) strategy has been choosen. For that purpose a 6x genome coverage of asparagus BAC library has been constructed. In order to get the target BAC clones linking to the M-locus, an AFLP-based DNA fingreprinting approach has been applied to screen a total number of 40992 BAC clones (about 2,6x genome coverage). Using 9 selected AFLP-primer combinations in total 13 different single BAC clones could be identified. This revealed 1,4 BAC clones per primer combinations, or lower than expected number 2,6 single BAC clones per primer combination. However the result indicated the feasibility of AFLP-based DNA-fingerprinting to be used in the screening of large insert library. Key words: Klon BAC, DNA-Fingerprinting, AFLP

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