| CHORI-259
Common Marmoset BAC Library |
 |
The CHORI-259 Common Marmoset (Callithrix jacchus, male)
BAC library has been constructed in Pieter De Jong's Laboratory
at the Children’s Hospital Oakland Research Institute,
BACPAC Resources, by Doriana Misceo and Dr. Baoli Zhu using
the cloning techniques developed in our laboratory (Osoegawa
et al., 1998). The marmoset sample for this library
was kindly provided by Dr.Suzette Tardif, Associate Director
of
the Southwest National Primate
Research Center in San Antonio TX. DNA was isolated
from kidney cells obtained from a male marmoset(SNPRC #17081).
Another sample from the same animal has been used to establish
a permanent cell line which can be obtained from the IPBIR
(Integrated Primate Biomaterials
and Information Resource). Agarose-embedded high molecular
weight DNA was partially digested with a combination of
EcoRI
restriction enzyme and EcoRI methylase. Size fractionated
DNA was ligated 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 528 384-well microtiter dishes and
also gridded onto 11 22x22cm nylon
high-density filter for screening by probe hybridization.
Each hybridization membrane represents over 18,000 distinct
BAC clones, stamped
in duplicate. BAC library construction was funded by NIH
grants HG01165-07SI and HG025323-01 as part of the NIH-funded
BAC Resource Network.
Provisional Data for Marmoset BAC library:
| Segment |
Cloning Vector |
DNA |
Restriction Enzyme |
Plate Numbers |
Total Plates |
Empty Wells |
Empty Wells (%) |
| 1 |
pTARBAC2.1 |
Adult Male kidney |
EcoRI |
1-288 |
288 |
1,506 |
1.4 |
| 2 |
pTARBAC2.1 |
Adult Male kidney |
EcoRI |
289-528 |
240 |
1,071 |
1.2 |
| Total Library |
|
|
|
1-528 |
569 |
2,577 |
1.3 |
| Segment |
Non-insert Clones (%) |
Non-insert clones |
Non-recombinant clones (BAC vector) |
Recombinant Clones |
Average Insert size (Kbp) |
Redundancy (Genome size:3.43 Gb) |
| 1 |
<1% |
N/A |
N/A |
N/A |
170 |
N/A |
| 2 |
|
N/A |
N/A |
N/A |
170 |
N/A |
| Total Library |
<1% |
N/A |
N/A |
N/A |
170 |
N/A |
click here for a legend of
the previous tables.
The CHORI-259 clone average insert size has been determined
by Pulsed Field Gel Electrophoresis. Clone size distribution
will be plotted as soon as available.
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
($11,520) or per segment
($5,760 for segment 1 and $4800 for segment 2).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).
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