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

Published online by Cambridge University Press:  29 November 2008

Pietro Piffanelli ,
Alberto D. Vilarinhos ,
Jan Safar ,
Xavier Sabau  and
Jaroslav Dolezel
Pietro Piffanelli
Affiliation:
Cirad-Bios, UMR DAP, av. Agropolis, TA A-96 / 03, 34398 Montpellier Cedex 5, France Parco Tecnol. Padano, Via Einstein, 26900 Lodi, Italy
Alberto D. Vilarinhos
Affiliation:
EMBRAPA Cassava Trop. Fruit Crop., Embrapa Str., Cruz das Almas, BA, 44380-000, Brazil
Jan Safar
Affiliation:
Lab. Mol. Cytogenet. Cytom., Inst. Exp. Bot., Sokolovska 6, CZ-77200 Olomouc, Czech Republic
Xavier Sabau
Affiliation:
Cirad-Bios, UMR DAP, av. Agropolis, TA A-96 / 03, 34398 Montpellier Cedex 5, France
Jaroslav Dolezel
Affiliation:
Lab. Mol. Cytogenet. Cytom., Inst. Exp. Bot., Sokolovska 6, CZ-77200 Olomouc, Czech Republic
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Abstract

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.

Keywords

FranceMusa sp.methodselectrophoresisDNAgenetic markersgenetic mapsFranceMusa sp.méthodeélectrophorèseADNmarqueur génétiquecarte génétique
Type
Research Article
Information
Fruits , Volume 63 , Issue 6 , November 2008 , pp. 375 - 379
Copyright
© CIRAD, EDP Sciences, 2008

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References

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