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 Home > Resources > Libraries
  CHORI-241 Equine BAC Library

  
  Background information

The CHORI-241 Horse (Equus caballus) BAC library has been generated by Baoli Zhu, in Pieter de Jong's laboratory at BACPAC Resources, Children's Hospital Oakland Research Institute. The library preparation followed the general cloning approach developed in our laboratory Osoegawa et al., 1998 . DNA was obtained from a blood sample provided by Dr. Doug Antczak of the Equine Genetic Center, James A. Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University.  The donor of this sample was a thoroughbred male horse by the name "Bravo", a half-brother of the female "Twilight" used for horse genome sequencing as part of the NIH horse genome project. A news release related to the project and the horses involved in the "BAC library" and "Horse Genome" Projects can be found in the online archive of The Cornell Daily Sun (http://cornellsun.com/node/21536). High molecular weight DNA was prepared by embedding peripheral blood neutrophil cells in an agarose matrix. (done by Dr. Helen Bird at Cornell). The library has been constructed by using partially digested DNA with EcoRI plus EcoRI methylase. Size fractionated DNA was cloned into the pTARBAC2.1 vector between the EcoRI sites. The ligation products were transformed into DH10B eletrocompetent cells (BRL Life Technologies). The library has been arrayed into 384-well microtiter dishes and also gridded onto a 22x22cm nylon high-density filter for screening by probe hybridization.

Segments

Cloning Vector

DNA

Restriction
enzyme

Plate Numbers

Total Plates

Empty Wells

Empty Wells (%)

1

pTARBAC2.1

Male Neutrophils

EcoRI

1-192

192

1,622

2.2%

2

pTARBAC2.1

Male Neutrophils

EcoRI

193-528

336

2,838

2.2%

Total Library

     

1-528

528

4,460

2.2%

 

Segments

Non-insert Clones (%)

Non-insert clones

Non-Recombinant Clones (pUC)

Recombinant Clones

Average Insert Size


Redundancy
(Genome size:
3.0 Mb)

1

2.1%

Approx. 1,514

552

Approx. 70,040

171Kbp

4.0

2

3.9%

Approx. 4,921

653

Approx. 120,612

171Kbp

6.9

Total Library

3.3%

Approx. 6,435

1,205

Approx. 190,652

171Kbp

10.9

 

The total library should represent approximately 11.8-fold total genomic representation.

Data on the CHORI-241 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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