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 Home > Resources > Libraries
   CHORI-226: fruit fly (Drosophila pseudoobscura MV2-25) BAC Library   

  Background information


The CHORI-226 library has been constructed by Chung-Li Shu and Kazutoyo Osoegawa in Pieter de Jong's laboratory, BACPAC Resources, Children's Hospital Oakland Research Institute. Agarose-embedded genomic DNA was prepared from Drosophila pseudoobscura MV2-25 strain by Dr. Stephen Richards (Human Genome Sequencing Center Baylor College of Medicine). To create the BAC clones, genomic DNA was partially digested with a combination of EcoRI and EcoRI Methylase. The restriction fragments were size-selected using pulsed-field gel electrophoresis and then ligated into the pTARBAC2.1 vector between the EcoRI sites. The ligation products were transformed into DH10B (T1 phage-resistant) electro-competent cells (Invitrogen). The library has been arrayed into 384-well microtiter dishes and has also been gridded onto 2 22x22cm nylon high-density filter for screening by probe hybridization. Each hybridization membrane represents about 18,000 distinct BAC clones, each represented in duplicate. BAC library construction was funded as part of the NIH-funded BAC Resource Network by a grant from NIH (#HG025323-02).

Strain information: The MV2-25 strain was collected by Dr. Wyatt Anderson (Department of Genetics, University of Georgia) in Mesa Verde CO in 1996. A population cage from this collection was set up from 8 isofemale lines on 7/8/1997. After approximately 50 generations within the population cage, 25 inbred lines were set up using virgin males and females from a single pair of parents. The DNA is derived from 25th line set up. Every generation, one brother and one sister (both virgins) were collected and crossed to each other. This inbreeding procedure was carried out for 14 generations. The species has been verified by inspection of the chromosomes cytologically by Dr. Anderson, and confirmed to be Drosophila pseudoobscura. Additionally inspection of the chromosomes confirmed the strain does not contain the arrowhead inversion, which was likely to cause problems in the sequencing of the first D. pseudoobscura strain.




Provisional data for CHORI-226 Drosophila pseudoobscura MV2-25 BAC Library:

Segment

1

All

Vector

pTARBAC2.1

pTARBAC2.1

Restriction Enzyme

EcoRI/EcoRI Methylase

EcoRI/EcoRI Methylase

DNA Source

Adult fly

Adult fly

Plate Numbers

1-96

1-96

Plate Count

96

96

Empty Wells

N/A

N/A

Non-Recombinant Clones

0 (0%)

0 (0%)

Non-Insert Clones

Approx. 402 (2/183, 1.09%)

Approx. 402 (2/183, 1.09%)

Recombinant Clones

36462

36462

Average Insert Size

125 Kbp

125 Kbp

Genomic Coverage

N/A

N/A



Data on the CHORI-226 clone insert size distribution 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 laboratory and data will be updated on our web page periodically.

Please contact Pieter J. de Jong for questions related to this library.


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 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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