Construction
of
a
deep
-
coverage
BAC
library
from
Lycopersicon
esculentum
cv
.
Heinz
1706
Budiman,
M
.
A
.
,
Frisch,
D
.
A
.
,
and
Wing,
R
.
A
.
Clemson
University
Genomics
Institute
(CUGI),
Clemson
University,
100
Jordan
Hall
BOX
345780,
Clemson,
SC
29634
-
5708
Large
insert
genomic
DNA
libraries
are
essential
for
many
genome
-
based
applications
from
positional
cloning
to
genome
sequencing
.
We
have
recently
completed
the
construction
of
a
deep
-
coverage
BAC
library
of
the
cultivated
tomato
-
Lycopersicon
esculentum
cv
.
Heinz
1706
.
Heinz
1706
was
selected
because
it
is
a
recurrent
parent
of
several
NILs
developed
by
Philouze
et
al
.
(1991)
and
is
the
jointed
parent
of
a
mapping
population
we
developed
to
map
jointless
-
2
.
Here
we
report
a
brief
description
of
the
library
and
how
it
will
be
provided
to
the
plant
community
.
Tomato
BAC
library
construction
and
characterization
The
BAC
library
was
constructed
by
size
selecting
Hind
III
partially
digested
tomato
genomic
DNA
and
ligating
with
pBeloBAC11
(kindly
provided
by
H
.
Shizuya,
Caltech),
followed
by
electroporation
into
E
.
coli
.
White
recombinant
clones
were
robotically
picked
and
arrayed
into
336
384
-
well
microtiter
plates
using
the
Q
-
BOT
system
(Genetix,
UK)
.
The
library
contains
129,000
clones
with
an
average
insert
size
of
117
.
5
kb
based
on
a
random
sampling
of
498
BAC
clones
(Figure
1)
.
Based
on
a
haploid
genome
size
of
tomato
of
930
Mbp/C
(Arumuganathan
and
Earle,
1991),
the
BAC
library
represents
approximately
15
genome
equivalents
and
thus
over
a
99
%
probability
of
recovering
any
specific
sequence
of
interest
.
0
2
4
6
8
10
12
14
16
18
20
Insert
size
range
(kb
)
Figure
1
.
Insert
size
distribution
of
Lycopersicon
esculentum
BAC
library
based
upon
the
analysis
of
498
random
BAC
clones
.
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