Adeno-Quest™

Adenoviral recombinants can be generated by in vivo homologous recombination in QBI-293A cells. Insertion of DNA by homologous recombination is the most efficient way of introducing a gene into an AdV because the genome is too large (36 kb) to be easily manipulated. Briefly, your cDNA is cloned into a transfer vector (step 1), then the transfer vector, containing the cloned gene in a transcription cassette adjacent to a homologous recombination sequence, is co-transfected into QBI-293A cells with a large part of the Ad5 genome. It is engineered so that the product of in vivo recombination between the two DNA molecules yields recombinant infectious viruses (step 2).

The Adeno-Quest™ kit comes in a complete package format containing all the principal components and controls for construction of 5 recombinant viruses. Each Adeno-Quest™ expression kit in-cludes all the basic components listed on page 20 plus your choice of one transfer vector.

On the following pages you will find a description of the reagents provided in the Adeno-Quest™ along with a transfer vector selection chart.

The Adeno-Quest™ kit comes in a complete package format containing all the principal components and controls for construction of 5 recombinant viruses. Each Adeno-Quest™ expression kit includes all the basic components listed below plus your choice of one transfer vector.

Description of Plasmids and Kit Reagents

Adeno-Quest™basic kit components

Cat No.	Product	Description	Quantity	Storage
AES0501	QBI-viral DNA	Ad5.CMV- LacZE1/E3 viral DNA Cla I cut	25µg (500ng/µl; 50µl)	-20°C
AES0507	QBI-Transf.	Ad5.CMV- LacZE1/E3 uncut DNA	2.5µg (50ng/µl; 50µl) (in 0.1 x sterile TE)	-20°C
AES0503	QBI-293A cells	Frozen 293 cell line	1ml (1 X 106 cells/ml)	-150°C
	QBI-Infect Ad5.CMV-LacZE1/E3	Viral particles of (in complete DMEM)	1ml (›1000 PFU/ml)	-80°C
	CaCl2 2M	Transfection reagent	0.5ml	-20°C
	TE 0.1x	Transfection reagent	0.5ml	-20°C
	HBS 2x	Transfection reagent	3 x 1ml	-20°C
	(see above)	Choice of transfer vector	25µg (500ng/ml; 50ml)	-20°C

Storage
-20°C to -150°C

Adeno-Quest™Transfer Vector selection chart

* MLP: Major Late Promoter of Adenovirus