|
RPCI-104: Schizosacharomyces pombe BAC Library |
 |
The RPCI-104 BAC Library has been constructed in our laboratory. Genomic DNA was purchased from
BIORAD (Cat# 170-3633) and partially digested with a combination
of EcoRI and EcoRI Methylase. Size selected DNA was cloned
into the pBACe3.6 vector between
the EcoRI sites. The ligation products were transformed into
DH10B electro-competent cells (BRL LIFE TECHNOLOGIES). The
library has been arrayed into 384-well microtiter dishes and
also gridded onto 11 x 7cm nylon high-density filters for
screening by probe hybridization. Each hybridization membrane
represents over 3,000 distinct Schizosaccharomyces pombe BAC
clones, stamped in duplicate. Library construction was supported
by sub-contracts from a grant awarded to Dr.
Kazuei Mita in The National Institute of Radiological
Sciences. The RPCI-104 Schizosaccharomyces pombe BAC Library
| Segment |
| DNA |
Restriction Enzyme |
Plate Numbers |
Total Plates |
Empty Wells |
Empty Wells (%) |
| 1 |
pBACe3.6 |
S.pombe 972H minus (ATCC 24843) |
EcoRI |
1-8 |
8 |
16 |
0.5 |
| Segment |
Non-insert Clones (%) |
Non-insert clones |
|
Recombinant Clones |
Insert size (Average) |
Redundancy (Genome
size:
15 Mb) |
| 1 |
1.2 |
Approx. 37 |
1 |
3018 |
166Kbp |
33 |
click here for a legend of the previous tables.
This library should represent approximately 33-fold total genomic representation.
Data on the RPCI-104 clone
average insert size has been determined by Pulsed Field
Gel Electrophoresis. Clone size distribution has been plotted
graphically. Complete
arrayed libraries are not generally available to individual
users, except for centers planning to establish genome-wide
annotation of the BAC library. Individual users may access
the library by ordering BAC clones as LB stab cultures. BAC
clone names should be known and can either be obtained from
public databases or by screening the library using a high-density
colony filter. The BAC names should comply with the NCBI
recommended clone nomenclature format. BAC Clones
identified after screening can be ordered through our online
ordering system or by Fax and require establisment of an account.
Please refer to ordering instructions on the BACPAC
homepage for procedures and costs. Academic and commercial
users interested in a copy of the BAC library should contact
Pieter J. de Jong ([email protected], fax: (510) 450-7924).
For hybridization membranes , please
contact BACPAC Resources ([email protected]).
Kazuei Mita, Ph.D.
(Present address)
Department of Genome Research
Laboratory of Insect Genome
National Institute of Agrobiological Sciences
1-2 Oowashi Thukuba-shi Ibaraki Japan
Tel: 0298-38-6120
Fax: 0298-38-6121
E-mail: [email protected]
|
