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CHORI-302: Amphioxus (Branchiostoma floridae) BAC Library |
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The CHORI-302 BAC 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 sperm of a Branchiostoma floridae which
was obtained by Dr. Linda Holland (Scripps Institution of
Oceanography) in Florida. 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 three 22x22cm nylon high-density
filter for screening by probe hybridization. The first
and second hybridization membrane represents about 18,000
distinct BAC clones, and the third filter contains about
11,000 clones, each represented in duplicate. Library characterization
was performed by Teresa Ren and Kazutoyo Osoegawa. Library
construction was supported by a sub-contract from a grant
(NSF IBN 0208138) awarded to Dr. Linda Holland. The library
is being characterized in collaboration with Drs. Linda Holland
and Jeremy Gibson-Brown, who have supplied us with a long
list of interesting genes to be screened.
| Cloning Vector |
pTARBAC2.1 |
| DNA Source |
Sperm |
| Restriction Enzyme |
EcoRI |
| Plate Range |
1-127 |
| Total 384-Well Dishes |
127 |
| Empty Wells (%) |
367 (0.8%) |
| Non-insert Clones (%) |
2/381 (0.5%) |
| Vector Clones (%) |
19 |
| Recombinant Clones |
>48,000 |
| Insert size (Average) |
142 Kbp |
| Library Redundancy (400 Mbp genome) |
~17 fold |
| Standard Deviation |
37 kb |
click here for a legend of
the previous tables.
Data on the CHORI-302 clone average
insert size has been determined by Pulsed Field Gel Electrophoresis.
Clone size distribution has been plotted graphically.
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 recipient's account number.
Organized general distribution of clones and screening reagents remains the exclusive activity of BACPAC Resources, unless
this service has ceased to exist. All library copies and
individual clones should be maintainedusing the recommended
nomenclature guidelines as posted on
the NCBI
web site.
When the scientific results obtained from use of the CHORI-302
library are published or otherwise disseminated, the scientific
creators and the origin of the BAC library should be acknowledged.
A recommended reference will be posted on this page as soon
as it is available. Otherwise, please contact Pieter
de Jong. This is important to permit us to measure the
impact of our scientific contributions and will facilitate
continued funding of similar resources.
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 ([email protected], fax: (510) 450-7924). For
hybridization membranes, please contact BACPAC
Resources ([email protected]).
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