| CHORI-106
Brugia malayi BAC Library |
 |
The CHORI-106 (BAC)
library has been constructed by Chung-Li Shu and Kazutoyo
Osoegawa in Pieter de Jong's laboratory, BACPAC Resources,
Children's Hospital Oakland Research Institute. Agarose-embedded
genomic DNA was prepared from Brugia malayi (TRS strain) by
Dr. Jeremy Foster (New England Boilabs). To create the BAC
clones, genomic DNA was MboI-partially digested. The restriction
fragments were size-selected using pulsed-field gel electrophoresis
and then ligated into the pTARBAC1.3 vector between the BamHI
sites. The ligation products were transformed into DH10B (T1
phage-resistant) electro-competent cells (Invitrogen). The
CHORI-106 is comprised of 4,500 clones (plates 1-12) with
an average insert size of 95 kb and 4,500 clones (plates 13-24)
with an average insert size of 138 kb. The total of 9,000
clones from both library segments provides ~9.4X haploid genome
(110 Mb) coverage. The arrayed clones have been gridded together
with CHORI-1106 fosmid library
onto a 22x22cm nylon high-density filter
for screening by probe hybridization. Each hybridization membrane
represents about 9,000 distinct BAC and 9,000 fosmid clones,
each represented in duplicate. Library construction was supported
by a sub-contract from a grant (NIH/NIAID U01-AI50903-02)
awarded to Dr. Elodie Ghedin at The Institute for Genomic
Research (TIGR).
Provisional
data for CHORI-106 Brugia malayi BAC library:
| Segment |
Cloning Vector |
DNA |
Restriction enzyme |
Plate Number |
Total Plates |
Empty Wells |
Empty Wells (%) |
| 1 |
pTARBAC1.3 |
TRS |
MboI |
1-12 |
12 |
135 |
2.9 |
| 2 |
pTARBAC1.3 |
TRS |
MboI |
13-24 |
12 |
136 |
2.9 |
| Total Library |
|
|
|
24 |
24 |
271 |
2.9 |
| Segment |
Non-insert clones
(%) |
Non-insert clones
|
Non-Recombinant
Clones (pUC) |
Recombinant clones |
Average Insert
Size |
Redundancy (Genome
Size: 110 Mb) |
| 1 |
1.7 |
Approx. 76 |
N/A |
Approx. 4,400 |
95 kbp |
3.8 |
| 2 |
0.6 |
Approx. 27 |
N/A |
Approx. 4,450 |
138 kbp |
5.6 |
| Total Library |
1.2 |
Approx. 103 |
N/A |
Approx. 8,850 |
117 kbp |
9.4 |
Data on the CHORI-106
clone average insert size has been determined by Pulsed
Field Gel Electrophoresis. Clone size distribution has been
plotted graphically. While analyzing clones using pulse-field
electrophoresis to determine the average insert size, non-insert
clones containing a small deleted vector fragment consistent
with sucrose resistance were observed. Further in depth characterization
of the library is on going in our lab and data will be updated
on our web page periodically.
Please direct questions
concerning this library to either Pieter
J. de Jong or Kazutoyo
Osoegawa.
| Ordering &
Pricing information |
The library is
available in several formats. Individual clones, and high-density
hybridization filters are
obtainable. For ordering and shipping details, please view
the ordering and pricing
information page.
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).
N/A
|
