| CHORI-253
Sumatran Orangutan BAC Library
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The CHORI-253 Sumatran Orangutan BAC library has been constructed
by Michael Nefedov in Pieter de Jong’s laboratory at
BACPAC Resources, Children’s Hospital Oakland Research
Institute. The preparation of the library followed the cloning
approach developed in our laboratory (Osoegawa
et al., 1998). Evan Eichler (Department of Genetics, Case
Western Reserve University) facilitated contacts with Steve
Taylor (Louisville Kentucky Zoo) to obtain a blood sample
from a male Sumatran orangutan ("Segundo"). DNA
was isolated from white blood cells by embedding the cells
in agarose. Agarose embedded DNA was partially digested with
a combination of EcoRI restriction enzyme and EcoRI Methylase
and the fragments were size-fractionated by pulsed-field gel
electrophoresis. DNA fragments from the appropriate size fraction
were cloned into the pTARBAC2.1 vector between the EcoRI sites.
The ligation products were then transformed into DH10B (T1
resistant) electro-competent cells (Invitrogen). The library
has been arrayed into 384-well microtiter dishes and also
gridded onto eleven 22x22cm nylon high-density
filters for screening by probe hybridization. Each hybridization
membrane represents over 18,000 distinct Orangutan BAC clones,
stamped in duplicate. Library characterization was performed
by Qing Cao and Kazutoyo Osoegawa. This work was funded by
NIH grant HG01165-06.
| Segment |
Cloning Vector |
DNA |
Restriction enzyme |
Plate Number |
Total Plates |
Empty Wells |
Empty Wells (%) |
| 1 |
pTARBAC2.1 |
Blood |
EcoRI |
1-288 |
288 |
2,126 |
1.9 |
| 2 |
pTARBAC2.1 |
Blood |
EcoRI |
289-528 |
240 |
2,096 |
2.3 |
| Total Library |
|
|
|
|
528 |
4,222 |
2.1 |
| Segment |
Non-insert clones (%) |
Non-insert clones (deleted vector) |
Non-Recombinant Clones (vector) |
Recombinant clones |
Insert Size (Average) |
Redundancy (Genome Size: 3000 Mb) |
| 1 |
0 |
0 |
0 |
108,000 |
177 Kbp |
6.4 |
| 2 |
0.4 |
~400 |
0 |
90,000 |
178 Kbp |
5.3 |
| Total library |
0.2 |
~400 |
0 |
198,000 |
177 Kbp |
11.7 |
Click here
for a legend of the previous tables.
Data on the CHORI-253 clone average insert size has been determined
by Pulsed Field Gel Electrophoresis. Clone
size distribution has been plotted graphically.
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
($10,560)
or per segment
($5,760 for segment 1 and $4,800 for segment 2). Shipping
will be done by Federal Express and charged to recipient's
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).
Please contact Pieter
J. de Jong or Kazutoyo
Osoegawa for questions related to this library.
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