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 Home > Resources > Libraries
  CHORI-223 Drosophila Melanogaster BAC Library

  
  Background information

The CHORI-223 Drosophila melanogaster BAC library has been constructed in our laboratory by Kazutoyo Osoegawa in collaboration with the Berkeley Drosophila Genome Project headed by Gerald M. Rubin and co-directed by Susan E. Celniker at Lawrence Berkeley National Laboratory. High-molecular-weight DNA from an isogenic y1; cn1 bw1 sp1 strain (B.J. Brizuela et al., 1994, Genetics 137: 803; Bloomington stock number 2057) was prepared by Roger A. Hoskins. The agarose embedded DNA was sheared by repeating freezing and thawing. The fragmented ends were polished by subsequent treatments with Mung bean nuclease and T4 DNA polymerase, respectively. The polished ends were ligated to the blunt-end side of an adapter which has a 3'overhang. Size fractionated DNA was cloned into the pTARBAC6 vector between the BstXI sites. The ligation products were transformed into DH10B electrocompetent 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. The hybridization membrane represents over 18,000 distinct D. melanogaster BAC clones, stamped in duplicate.


Provisional data for CHORI-223 Drosophila melanogaster BAC library:

Segments Cloning Vector DNA Restriction
enzyme
Plate Numbers Total Plates Empty Wells Empty Wells (%)
1 pTARBAC6 Adult fly sheared 1-48 48 258 1.4

 

Segments
Non-insert Clones (%)
Non-insert clones
Non-Recombinant Clones (pUC)
Recombinant Clones
Average Insert Size

Redundancy
(Genome size:
120 Mb euchromatic portion)
1
1.0
182
not identified yet
17,992
48 Kbp
7.2

 

The total library should represent approximately 7-fold total genomic representation.

Data on the CHORI-223 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.

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

Osoegawa K, Vessere GM, Li Shu C, Hoskins RA, Abad JP, de Pablos B, Villasante A, de Jong PJ. BAC clones generated from sheared DNA. Genomics 2007 89:291-9.

 

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