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 Home > Resources > Libraries
  CHORI-211 Zebrafish BAC Library

  
  Background information

The CHORI-211 BAC library has been constructed by Chung Li Shu and Kazutoyo Osoegawa in our laboratory. Genomic DNA was isolated from testis of 6 males by Dr. Gerd-Joerg Rauch and partially digested with a combination of EcoRI and EcoRI Methylase. Size selected DNA was cloned into the pTARBAC2.1 vector between the EcoRI sites. The ligation products were transformed into DH10B electro-competent cells (BRL LIFE TECHNOLOGIES). The library has been arrayed into 384-well microtiter dishes and also gridded onto 6 22x22cm nylon high-density filters for screening by probe hybridization. Each hybridization membrane represents over 18,000 distinct zebrafish BAC clones, stamped in duplicate. Library construction was supported by sub-contracts from a grant awarded to Dr. Robert Geisler at The Max-Planck-Institut fuer Entwicklungsbiologie.

The strain name of zebrafish is Tue (Tuebingen).
The characteristics of this fish line are described in the following paper:
Rauch, G.-J., Granato, M. and Haffter, P. (1997). A polymorphic zebrafish line for genetic mapping using SSLPs on high-percentage agarose gels. Technical Tips Online T01208.


Provisonal data for CHORI-211 Zebrafish BAC Library:

Segment
DNA
Restriction Enzyme
Plate Numbers
Total Plates
Empty Wells
Empty Wells (%)
1
pTARBAC2.1
Testis from 6 males
EcoRI
1-144
144
1,314
2.4
2
pTARBAC2.1
Testis from 6 males
EcoRI
144-288
144
1,278
2.3

 

Segment
Non-insert Clones (%)
Non-insert clones
Recombinant clones
Insert size (Average)
Redundancy (Genome size:
1,700 Mb)
1
1.2
approx. 648
0
53,334
171 Kbp
5.4
2
2.6
approx. 1404
41
52,573
160 Kbp
5.0

Data on the CHORI-211 clone average insert size of Segment 1 and Segment 2 have been determined by Pulsed Field Gel Electrophoresis. Clone size distribution for each segment 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.

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

  Ordering & Pricing information

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).

  Reference

N/A

 

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