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ARS Home » Plains Area » Miles City, Montana » Livestock and Range Research Laboratory » Research » Publications at this Location » Publication #120757
Title: CONSTRUCTION OF NEW MAIZE, BOVINE, EQUINE AND ZEBRAFISH BAC LIBRARIES

Author
item OSOEGAWA, KAZUTOYO - CHILDREN'S HOSP OAKLAND
item LI SHU, CHUNG - CHILDREN'S HOSP OAKLAND
item ZHU, BAOLI - CHILDREN'S HOSP OAKLAND
item GARDNER, JACK - UNIVERSITY OF MISSOURI
item Geary, Thomas
item ANTEZAK, DOUG - CORNELL UNIVERSITY
item RAUCH, GERD-JOERG - MAX-PLANCK-INSTITUTE
item CATANESE, JOSEPH - CHILDREN'S HOSP OAKLAND
item DE JONG, PIETER - CHILDREN'S HOSP OAKLAND

Submitted to: Plant and Animal Genome Conference Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: 2/20/2001
Publication Date: 2/20/2001
Citation: OSOEGAWA, K., LI SHU, C., ZHU, B., GARDNER, J., GEARY, T.W., ANTEZAK, D., RAUCH, G., CATANESE, J.J., DE JONG, P.J. CONSTRUCTION OF NEW MAIZE, BOVINE, EQUINE AND ZEBRAFISH BAC LIBRARIES. PLANT AND ANIMAL GENOME CONFERENCE PROCEEDINGS. 2001. Available: www.intl-pag.com/pag/9/abstracts.

Interpretive Summary: We have developed efficient cloning procedures for construction of BAC libraries by partial restriction digestion (Osoegawa et al., Genomics 1998, 52: 1-8). To support genomic research by the scientific community more than 30 BAC libraries for various organisms have been prepared in our laboratory and have been distributed. Maize genomic DNA was prepared from nuclei, was digested with EcoRI or MboI and then cloned into our pTARBAC vectors. About 107,000 clones from EcoRI-digested DNA have been arrayed into 384-well dishes. A random set of 295 clones was analyzed using pulsed-field gel electrophoresis revealing an average insert size of 163 kb, consistent with 6.9-fold genome redundancy of the EcoRI maize library. The arraying and characterization of the MboI maize library is underway. The maize library has been designated as CHORI-201, segment 1 (EcoRI) and segment 2 (MboI). Mammalian (bovine and equine) and zebrafish BAC libraries are currently under construction and will each be represented at 10-fold redundancy. Genomic DNA was prepared from leukocytes of a Line 1 Hereford Bull, from neutrophils of a horse or from zebrafish testis. The DNA, digested with MboI for bovine, and EcoRI for equine and zebrafish, is being cloned into our pTARBAC vectors. Information on our available BAC libraries and vectors is available through www.chori.org/bacpac. All BAC libraries have been constructed, either directly or indirectly, through support by grant funds from the USDA, NSF, NHGRI, US DOE and other agencies.

Technical Abstract: We have developed efficient cloning procedures for construction of BAC libraries by partial restriction digestion (Osoegawa et al., Genomics 1998, 52: 1-8). To support genomic research by the scientific community more than 30 BAC libraries for various organisms have been prepared in our laboratory and have been distributed. Maize genomic DNA was prepared from nuclei, was digested with EcoRI or MboI and then cloned into our pTARBAC vectors. About 107,000 clones from EcoRI-digested DNA have been arrayed into 384-well dishes. A random set of 295 clones was analyzed using pulsed-field gel electrophoresis revealing an average insert size of 163 kb, consistent with 6.9-fold genome redundancy of the EcoRI maize library. The arraying and characterization of the MboI maize library is underway. The maize library has been designated as CHORI-201, segment 1 (EcoRI) and segment 2 (MboI). Mammalian (bovine and equine) and zebrafish BAC libraries are currently under construction and will each be represented at 10-fold redundancy. Genomic DNA was prepared from leukocytes of a Line 1 Hereford Bull, from neutrophils of a horse or from zebrafish testis. The DNA, digested with MboI for bovine, and EcoRI for equine and zebrafish, is being cloned into our pTARBAC vectors. Information on our available BAC libraries and vectors is available through www.chori.org/bacpac. All BAC libraries have been constructed, either directly or indirectly, through support by grant funds from the USDA, NSF, NHGRI, US DOE and other agencies.