| CHORI-252
Vervet Monkey BAC Library
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The CHORI-252 vervet monkey 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). A blood sample was obtained from an adult
male vervet monkey (#1994-021) through Roel A. Ophoff (Center
for Neurobehavioral Genetics, UCLA) and Lynn A. Fairbanks
(Center for Primate Neuroethology, UCLA). DNA was isolated
from white blood cells by embedding the cells in agarose.
Agarose embedded high molecular weight DNA was partially digested
with a combination of EcoRI restriction enzyme and EcoRI Methylase.
The DNA fragments were size-fractionated by pulsed field electrophoresis
and 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) electrocompetent 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 vervet monkey BAC
clones, stamped in duplicate. Library characterization was
performed by Qing Cao and Kazutoyo Osoegawa. This work was
funded by NIH grant HG01165-06.
Note: This BAC library
has been used for large-scale physical mapping by BAC-end
sequencing and comparative alignment of the BAC-end Sequences
(BES) relative to the human and rhesis macaque genome assemblies.
One can search for BACs containing any "putative"
vervet genes by looking for those genes in the UCSC Human
Genome Browser displaying a custom track of the CHORI-252
BACs relative to the human genome. The BAC-end sequences were
obtained at McGill University, Montreal as funded by Genome
Quebec and Genome Canada. More information can be found at
websites displaying the Vervet Monkey Physical
Map Project and the Integrated
Vervet Monkey Genomic Resources. From the Physical
Map Project URL, one can upload all the mapped BAC-end Sequences
to the UCSC Human Genome Browser by following the recommendation
"You can go directly to
the UCSC Genome Browser here
" .
| Segment |
Cloning Vector |
DNA |
Restriction enzyme |
Plate Number |
Total Plates |
Empty Wells |
Empty Wells (%) |
| 1 |
pTARBAC2.1 |
Blood |
EcoRI |
1-288 |
288 |
1,236 |
1.1 |
| 2 |
pTARBAC2.1 |
Blood |
EcoRI |
289-528 |
240 |
726 |
0.8 |
| Total Library |
|
|
|
|
528 |
1,962 |
1 |
| Segment |
Non-insert Clones (%) |
Non-insert Clones (deleted vector) |
Non-recombinant Clones (vector) |
Recombinant Clones |
Insert Size (Average) |
Redundancy (Genome Size: 3,000 Mb) |
| 1 |
0.4 |
~ 400 |
13 |
110,000 |
160 Kbp |
5.9 |
| 2 |
0 |
~ 0 |
15 |
91,000 |
160 Kbp |
4.9 |
| Total library |
0.2 |
~ 400 |
28 |
.201,000 |
160 Kbp |
10.8 |
click here
for a legend of the previous tables.
Data on the CHORI-252 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.
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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