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CHORI-103: Schistosoma mansoni BAC Library |
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The CHORI-103 BAC
library has been constructed by Chung Li Shu and
Kazutoyo Osoegawa at BACPAC Resources, Children's Hospital
Oakland Research Institute. The NMRI strain was used for construction
of the library. The strain has been in continuous culture
since the 1950's. It has undergone a number of bottle necks
and is likely highly inbred. Genomic DNA was isolated from
the cercariae in agarose by Dr. Philip T. LoVerde and partially
digested with MboI. Size selected DNA was cloned into the
pTARBAC1.3 vector between the BamHI sites. The ligation products
were transformed into DH10B T1 phage resistant electrocompetent
cells (BRL Life Technologies). The library has been arrayed
into 384-well microtiter dishes and also gridded onto two
22x22cm nylon high-density filters (one for each library
segment) for screening by probe hybridization. Each hybridization
membrane represents over 18,000 distinct Schistosoma mansoni
BAC clones, stamped in duplicate. Library construction was
supported through a sub-contract from a grant (U01 AI48828
from NIAID (NIH)) awarded to Dr. Najib M. El-Sayed at The
Institute for Genomic Research (TIGR).
Provisional
data for CHORI-103 Schistoma mansoni BAC library:
| Segment |
Cloning Vector |
DNA |
Restriction enzyme |
Plate Numbers
| Total Plates |
Empty Wells |
Empty Wells (%)
| | 1 |
pTARBAC1.3 |
cercariae |
MboI |
1-48 |
48 |
485 |
2.6 |
| 2 |
pTARBAC1.3 |
cercariae |
MboI |
49-96 |
48 |
523 |
2.8 |
| Segment |
Non-insert clones (%) |
Non-insert clones |
Non-Recombinant Clones (pUC) |
recombinant clones |
Average Insert Size |
Genomic Coverage
(270Mb) |
| 1 |
4.5 |
Approx. 808 |
0 |
Approx. 17,139 |
140 Kbp |
8.9X |
| 2 |
1.9 |
Approx. 340 |
0 |
Approx. 17,569 |
150 Kbp |
9.8X |
Data
on the CHORI-103 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 .
| 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 ([email protected]),
fax: (510) 450-7924).
Dr.
Philip T. LoVerde ([email protected])
University at Buffalo
138 Farber Hall
School of Medicine and Biomedical Sciences, 3435 Main Street
Buffalo, New York 14214
Dr. Najib M. El-Sayed
([email protected])
Assistant Investigator
Department of Eukaryotic Genomics
The Institute for Genomic Research
9712 Medical Center Drive, Rockville, MD 20850
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