AdEasy system is used to rapidly generate recombinant adenovirus
without the need for time consuming plaque purification. Developed by
T.C He et al (12), the AdEasy system exploits E. coli's robust,
efficient recombination machinery thereby avoiding restriction-ligation
involving the unwieldy (36kb) adenovirus genome. Bacterial and 293 cells
that are essential to the technique are included in every kit with each
kit providing enough reagents to generate up to 5 recombinant adenoviruses.
The highly detailed user manual is an invaluable resource for beginners
as well as those experienced in this field of work.
STEP 1 : The cDNA of interest is first cloned into a transfer vector.
STEP 2 : The resulting plasmid is linearized with Pme I and co-transformed into E. coli strain BJ5183 together with pAdEasy-1, the viral DNA plasmid. Recombinants are selected with kanamycin and screened by restriction enzyme analysis.
STEP 3 : The recombinant adenoviral construct is then cleaved with Pac I to expose its ITR (Inverted Terminal Repeats) and transfected into QBI-293A cells to produce viral particles.
The AdEasy kit comes in a complete package format containing all the principal components and controls for the construction of 5 recombinant viruses. Each AdEasy system kit includes all the components listed below plus your choice of one transfer vector.
Contents of the AdEasy Kit
-20°C to -150°C