| 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)
The library is available for academic users by reimbursement of duplication
costs (labor & materials), resulting in a price of $20 per 384-well dish.
Dishes can only be obtained per complete library
($2,880).Individual clones identified after screening can be obtained as stab cultures
at a cost of $25 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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