Construction of bacterial artificial chromosome (BAC) libraries of banana (Musa acuminata and Musa balbisiana).

Introduction. This protocol applies to the characterization of genome structure and evolution of M. acuminata and M. balbisiana genomes; construction of physical maps around markers of interest and QTLs; comparative genomic studies with other monocot species; and study of banana streak virus (BSV) integration patterns in banana nuclear genomes. The principle, key advantages, starting plant material, time required and expected results are presented. Materials and methods. This part describes the required materials, and the protocols for isolation and lysis of banana nuclei; high-molecular-weight (HMW) DNA Hind III digestion and pulsed-field gel electrophoresis (PFGE) analysis; large-scale HMW DNA Hind III digestion and ligation to the pINDIGOBAC-5 vector; and analysis of recombinant BAC clones. It mentions the main problems which can occur. Results. The described protocols will enable the efficient construction of BAC libraries from Musa species. These libraries can be used to construct physical maps and select specific genomic regions to be sequenced.

Bibliographic Details

Main Authors:	PIFFANELLI, P., VILARINHOS, A. D., SAFAR, J., SABAU, X., DOLEZEL, J.
Other Authors:	Pietro Piffanelli, CIRAD-BIOS/Parco Tecnologico Padano; Alberto Duarte Vilarinhos, CNPMF; Jan Safar, Institute of Experimental Botany; Xavier Sabau, CIRAD-BIOS; Jaroslav Dolezel, Institute of Experimental Botany.
Format:	Artigo de periódico biblioteca
Language:	Ingles English
Published:	2009-02-26
Subjects:	Method, Genetic marker, Genetic map, DNA, Musa sp, electrophoresis,
Online Access:	http://www.alice.cnptia.embrapa.br/alice/handle/doc/655566
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