Virus Clone Isolation by Plaque Assay

It is good virological procedure to purify a recombinant virus by plaque assays, even if the virus was generated from a plasmid or cosmid isolated from a single E. coli clone. A recombinant adenovirus can prove unstable if the expression cassette it carries exceeds the maximal transgene capacity of the vector, or if it encodes a protein that is toxic for the helper cells or interferes with viral replication.

Applications:

  • Isolation of viral clones during virus construction, prior to large-scale virus amplification
  • Isolation of clones of a virus that might reveal unstable
  • Isolation of clones from a virus suspension that contains replication-competent adenovirus (RCA)
Top

You provide:

  • A virus suspension
Top

The service includes:

  • Infection of monolayers of helper cells with a serial dilution of your virus – agar overlay
  • Isolation of 10 plaques
  • Small-scale amplification of each plaque
  • Confirmation of virus identity at the DNA level (restriction analysis, PCR or Southern blot) or protein expression (Western-blot, RT-PCR).
Top

Advantages of using our services:

  • Short turn-around
  • Reliability
  • Cost-effective
Top

Minimum time required:

  • 2 weeks
Top

An Important Note on Handling Adenovirus:

Adenovirus is an infectious respiratory virus and its use as a genetic vector requires the use of adequate containment equipment and practices. Biosafety Level 2 (BL2) is appropriate for most customers; however, for some, use of BL3 or higher level containment facility may be required. PLEASE NOTE, O.D.260 Inc. will not carry out projects that require BL3 (or above) containment.

Services | How to Proceed | Pricing

 

PO Box 534 Boise, ID 83701   |   Phone: (208) 345-7369   |   Fax: (208) 345-7569   |   info@od260.com