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AdEasy™ Adenoviral Vector System
Description of the method
 The
AdEasy™ system is fundamentally the same as the AdenoVator™
system. Both systems use the homologous recombination system in
E. coli cells (BJ5183), the same Ad5-backbone carrying plasmid
(pAdEasy-1), and the same cells for the amplification of the recombinant
adenovirus. The only difference between these two systems lies in
the transfer vectors available. The AdEasy™ and AdenoVator™systems
are 100% compatible.
Advantages
- No plaque purification required
- Uses the highly efficient homologous recombination apparatus
of E. coli
- Vector selection allows you to subclone your own expression
cassette, including your choice of promoter and polyA signal
- 100% compatible with the AdenoVator™ adenoviral vector
system
Cell Lines and Viruses
BJ5183 EC
BJ5183 EC are electrocompetent (EC) Escherichia coli cells
used in the AdenoVator™ and AdEasy™ systems for homologous
recombination. This strain is not suited for amplification of recombinant
plasmid. Genotype: endA sbcBC recBC galK
met thi-1 bioT hsdR (Strr).
DH5
EC
DH5
EC are electrocompetent Escherichia coli cells used in the
AdenoVator™ and AdEasy™ kits for amplification of recombinant
plasmid. This strain is not suited for homologous recombination.
Genotype: F- deoR recA1 endA1 hsdR17(rk-,
mk ) supE44  -
thi-1 gyrA96 relA1.
QBI-HEK 293A Cells
The QBI-HEK 293A cell line is a permanent line of primary human
embryonic kidney (HEK) cells transformed by sheared human Ad5 DNA.
These cells contain the E1A and E1B Ad5 viral genes, which complement
the deletion of this essential region in the recombinant adenovirus.
We have selected a superior sub-clone of HEK 293 that adheres strongly
to plastic dishes and performs extremely well in plaque assay and
transfection experiments. All our adenoviral vector systems use
QBI-HEK 293A cells for replication and amplification.
QBI-Infect Viral Particles
The QBI-Infect is a first generation adenovirus expressing the
bacterial LacZ gene under the control of the immediate/early CMV
promoter (Ad5.CMVLacZ E1/E3).
This virus can be used to perfect adenovirus culture techniques
before producing recombinant virus. People inexperienced with virology
techniques can use QBI-Infect to prepare control plaques. Once
amplified, QBI-Infect Adenovirus can be used to test the infectivity
of cell lines or tissues other than QBI-HEK 293A cells or commonly
used eukaryotic cells. The success of DNA transfer can be easily
monitored with a substrate for  -galactosidase.
The QBI-Infect Viral Particles are included in all of our adenoviral
vector systems.
Kit Information
The basic kit contains the following components:
- BJ5183 cells
- DH5
cells
- pAdEasy-1
- QBI-Infect
- Transfection reagents (2M CaCl2, TE 0.1X and 2X HBS)
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Reagents provided |
| Cat # |
Basic Kit |
Transfer Vector |
QBI-HEK293A cells |
| AES1000 |
X |
none |
X |
| AES1000A |
X |
pShuttle and pShuttle-CMV |
X |
| AES1001 |
X |
none |
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| AES1001A |
X |
pShuttle and pShuttle-CMV |
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Products sold separately
| Cat # |
Description |
Volume |
| AES1010 |
pAdEasy-1, the 33.4 kb plasmid containing the E1/E3
Adenovirus serotype 5 genome |
2.5 µg |
| AES1020 |
pShuttle transfer vector |
25 µg |
| AES1021 |
pShuttle-CMV transfer vector |
25 µg |
| AES1009 |
Electrocompetent bacteria BJ5183 DH5 |
5 tubes each |
| AES1005K |
EC (Electrocompetent) BJ5183 cells |
5 X 80 µL |
| AES1007K |
EC (Electrocompetent) DH5 |
5 X 40 µL |
| AES0503 |
QBI-HEK 293A cells |
1 mL (1X 106 cells) |
| AES1015 |
Refill kit: pAdEasy-1 BJ5183 EC cells |
Kit |
| AES1016 |
Refill kit: pAdEasy-1 BJ5183 EC cells DH5 |
Kit |
To obtain additional contact or ordering information, click
here.
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