| CHORI-260
Turkey BAC Library
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The CHORI-260 Turkey 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 from a female turkey was
kindly provided by Dr. David Harry (Nicholas Turkey Breeding
Farms, Sonoma, CA). DNA was isolated from whole blood by embedding
a mixture of erythrocytes and lymphocytes in agarose. The
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) electro-competent cells (Invitrogen). The library
has been arrayed into 192 [384-well] microtiter dishes. For
screening by probe hybridization, four 22x22cm
nylon high-density colony filters have been prepared.
Each hybridization membrane represents over 18,000 distinct
BAC clones from 48 microtiter dishes, represented by duplicate
colonies. Library characterization (see below) was performed
by Qing Cao, Teresa Ren & Kazutoyo Osoegawa. This work
was funded by grant support from the USDA (#99-35205-8537).
| Cloning Vector |
pTARBAC2.1 |
| DNA Source |
Whole blood |
| Restriction Enzyme |
EcoRI |
| Plate range |
(1 - 192) |
| Total 384-well dishes |
192 |
| Empty wells |
1,500 (<2.1 %) |
| Non-insert clones |
2/182(<1.1%) |
| Vector clones |
0/73k (<<1%) |
| Recombinant Clones |
~ 71,000 (>96% of wells) |
| Insert Size Average |
~ 190 kb |
| Standard Deviation |
44 kb |
| Library Redundancy |
~11.1 fold |
Click here for a legend of
the previous tables.
Data on the CHORI-260 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 ($3,840). 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.
Please contact Pieter J.
de Jong or Kazutoyo
Osoegawa for questions related to this library.
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