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 Home > Resources > Libraries
  CHORI-301 Ciona savignyi BAC library

  

The CHORI-301 Ciona savignyi BAC library has been constructed in our laboratory by Kazutoyo Osoegawa in collaboration with Drs. Arend Sidow's and William C. Smith's laboratories. DNA was isolated from sperm. The library has been constructed using DNA which was sheared as part of the normal liquid DNA extraction procedure. 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 3 22x22cm nylon high-density filters for screening by probe hybridization. Each hybridization membrane represents about 18,000 distinct Ciona savignyi BAC clones, stamped in duplicate. Library construction was supported by sub-contracts from UC Santa Barbara through NSF grants awarded to Dr. William C Smith.


Segment Cloning Vector DNA

Restriction Enzyme

Plate Numbers Total Plates empty wells Empty Wells (%)
1 pTARBAC6 Sperm from 25 individuals Sheared 1-144 144 1021 1.8

 

Segment Non-insert clones (%) Non-insert clones Non-Recombinant Clones (pUC) Recombinant clones Insert size (average) Redundancy (Genome size: 180Mb)
1 1.0 543 No data 53,732 46 Kbp 13.7

click here for a legend of the previous tables.

The total library should represent approximately 14-fold total genomic representation.

The CHORI-301 clone average insert size has been determined by Pulsed Field Gel Electrophoresis. Clone size distribution has been plotted graphically.

The library is available as library plates, individual clones, and high-density hybridization sets (see Filter Availability)

Individual clones identified after screening can be obtained as stab cultures at a cost of $80 per clone in addition to a $35 set-up charge per shipment. Shipping will be done by Federal Express and charged to the recipients account number.

With respect to the scheduling of shipments of arrayed library copies, we would like to make you aware of our policy for array distribution.

Please visit the ordering information page if you want to place an order.

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). For hybridization membranes, please contact BACPAC Resources (BACPACorders@chori.org).

 

 


Dr. Arend Sidow (arend@stanford.edu)
Departments of Pathology and Genetics
SUMC R248B
300 Pasteur Drive
Stanford, CA 94305-5324

Dr. William C Smith (w_smith@lifesci.ucsb.edu)
MCD Biology
University of California
Santa Barbara, CA 93106

 

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