| CHORI-213
Three-spine Stickleback BAC Library
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The CHORI-213 Three-spine
Stickleback BAC library has been constructed by Baoli Zhu
in Pieter de Jong’s Laboratory at the Children’s
Hospital Oakland Research Institute using the cloning technique
developed in our laboratory (Osoegawa
et al., 1998). DNA was isolated from blood obtained from
Three-spine Sticklebacks by Dr.
Chris Amemiya Benaroya Research Institute at Virginia Mason).
Agarose embedded high-molecular-weight DNA was partially digested
with a combination of EcoRI restriction enzyme and EcoRI Methylase.
Size fractionated DNA was cloned into the pTARBAC2.1 vector
between the EcoRI sites. The ligation products were transformed
into DH10B (T1 resistant) electrocompetent cells (Invitrogen).
The library has been arrayed into 384-well microtiter dishes
and also gridded onto four 22x22cm nylon
high-density filters for screening by probe hybridization.
Each hybridization membrane represents over 18,000 distinct
stickleback BAC clones, stamped in duplicate. Library construction
was supported through a sub-contract from a grant awarded
to Dr. David Kingsley at Stanford University.
Provisional
data for CHORI-213 Three-spine Stickleback BAC Library:
| Cloning Vector |
DNA |
Restriction enzyme |
Plate Number |
Total Plates |
Empty Wells |
Empty Wells (%) |
| pTARBAC2.1 |
Blood |
EcoRI |
1-192 |
192 |
1,387 |
1.9 |
| Non-insert clones (%) |
Non-insert clones |
Non-Recombinant Clones (pUC) |
recombinant clones |
Insert Size (Average) |
Redundancy (Genome Size: 700Mb) |
| 1.3 |
Approx.988 |
264 |
Approx. 71,087 |
190 Kbp |
19.3 |
Data on the CHORI-213 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.
| Pricing &
Ordering 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).
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