Recently,
we developed methods that enable us to produce High Quality Nanoquantity
cDNA Libraries from such small amounts of tissue and cells that
material prepared by laser capture microdissection (LCM) or FACS
can be used. Like our standard libraries, these libraries are directionally
cloned into Express I and then transformed into T1 phage resistant
E. coli. We guarantee at least 2x105
primary clones with 87%
recombinant clones containing an average insert size of at least
800 bp, however, we generally produce libraries of 5x105
primary clones with >95% recombinant clones containing 1.0kb
average insert size.
We
will make nanoquantity libraries from customer supplied tissue (at
least 250 µg), cells (at least 25,000), total RNA ( 250
ng) or mRNA ( 5
ng). We provide quality controlled primary or amplified libraries.
Features list:
- High quality cDNA clones.
- Directionally cloned cDNA for expression, antibody screening,
subtraction and normalization.
- Large numbers of primary clones.
- Low vector background.
Quality testing: Percentage of recombinant clones
and average insert size determined by gel analysis of 24 clones
picked at random.
Shipment and storage conditions: Libraries will
be shipped in dry ice. Storage of libraries at -80°C is recommended.
Characteristics
of a cDNA Library Constructed From a Nanoquantity
Amount of HeLa mRNA |
mRNA
(ng) |
Total
# of primary clones |
Average
insert size (kb)* |
Percent
recombinants |
Insert
size range (kb)* |
100 |
7.2x107
|
1.20
|
100
|
0.5
- 3.2 |
50 |
1.5x107
|
1.35
|
100 |
0.6
- 3.5 |
10 |
1.7x106 |
1.00
|
100
|
0.4
- 2.9 |
5 |
9.0x105 |
1.01
|
96 |
0.4
- 3.0 |
2 |
2.0x105 |
0.74
|
94 |
0.2
- 2.3 |
1 |
4.5x104 |
0.92
|
94 |
0.2
- 3.6 |
*Average
insert size and insert size range determined by restriction enzyme
digestion of 24 clones picked at random from each library.
Click
here for Tissue and RNA Preparation suggestions |
|