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 Home > Resources > Libraries
  CHORI-240 Bovine BAC Library (Male)   
  Background information

 

The CHORI-240 BAC library has been constructed by Chung Li Shu and Kazutoyo Osoegawa in our laboratory. Genomic DNA was isolated from Hereford bull L1 Domino 99375 and was partially digested with MboI. Size selected DNA was cloned into the pTARBAC1.3 vector between the BamHI sites. The ligation products were transformed into DH10B electrocompetent cells (BRL Life Technologies) for segment 1 and DH10B T1 phage resistant electrocompetent cells (BRL Life Technologies) for segment 2, respectively. The library has been arrayed into 384-well microtiter dishes and also gridded onto 11 22x22cm nylon high-density filters for screening by probe hybridization. Each hybridization membrane represents over 18,000 distinct bovine BAC clones, stamped in duplicate. BAC library construction was funded by a grant from USDA (99-35205-8537).

Blood was collected from Hereford bull L1 Domino 99375 in Dr. Michael MacNeil's laboratory, USDA-ARS, Miles City, MT. For additional information with respect to the bovine strain, please contact Dr. Michael MacNeil by email (mike@larrl.ars.usda.gov).

Segment
DNA
Restriction Enzyme
Plate Numbers
Total Plates
Empty Wells
Empty Wells (%)
1
pTARBAC1.3
Blood
MboI
1-288
288
2,160
2.0
2
pTARBAC1.3
Blood
MboI
289-528
240
1,207
1.3
Total
1-528
528
3,367
1.7

 

Segment
Non-insert Clones (%)
Non-insert clones
Recombinant Clones
Insert size (Average)
Redundancy (Genome size:
3000 Mb)
1
5.7
Approx. 6,180
0
Approx. 102,252
180 Kbp
6.1
2
1.6
Approx. 1,455
15
Approx. 89,483
153 Kbp
4.6
Total
15
Approx. 191,735
167 Kbp
10.7


Data on the CHORI-240 clone average insert size has been determined by Pulsed Field Gel Electrophoresis. Clone size distribution has been plotted graphically. While analyzing clones using pulse-field electrophoresis to determine the average insert size, non-insert clones containing a small deleted vector fragment consistent with sucrose resistance were observed. Further in depth characterization of the library is on going in our lab and data will be updated on our web page periodically.

The library is available in several formats. Individual clones, and high-density hybridization filters are obtainable. For ordering and shipping details, please view the ordering and pricing information  page.

Academic and commercial users interested in a copy of the BAC library should contact Pieter J. de Jong ( pdejong@chori.org ), fax: (510) 450-7924).


Please direct questions concerning this library to either Pieter J. de Jong or Kazutoyo Osoegawa.

 

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