| CHORI-261
Chicken BAC Library |
 |
The CHORI-261 Chicken 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 whole blood sample from a chicken (jungle
fowl, female) was kindly provided by Jerry Dodgson (Michigan
State University). The animal (Jungle Fowl UCD001, Inbred
256 (female) used for the blood sample has also been used
for three additional BAC libraries prepared at Texas
A&M University and being used for the chicken
genome sequencing project at Washington
University in St.Louis. DNA was isolated from the 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 Teresa Ren, Qing Cao, & Kazutoyo
Osoegawa. This work was funded by grant support from the USDA
under the auspices of the USDA-CSREES National Animal Genome
Research Program (USDA Grant: #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 |
2,158 (-2.90%) |
| Non-insert clones |
0/190(<0.5%) |
| Vector clones |
1/73k (<<1%) |
| Recombinant Clones |
~ 73,000 (>97% of wells) |
| Insert Size Average |
~ 182 kb |
| Standard Deviation |
36 kb |
| Library Redundancy |
~11 fold |
Click here for a legend of
the previous tables.
Data on the CHORI-261
clone average insert size
has been determined by Pulsed Field Gel Electrophoresis. Clone
size distribution has been plotted graphically.
Hybridization filters can be
ordered only per complete set of 4 filter membranes (price
per set is 4 times $200). Shipping of arrayed library copies
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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