UNIT 6.8 Immunoscreening after Hybrid Selection and Translation
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Velan, B. 2001. Immunoscreening after Hybrid Selection and Translation. Current Protocols in Molecular Biology. 24:IV:6.8:6.8.1–6.8.5.
- Published Online: 1 MAY 2001
- Published Print: JAN 1991
In this procedure, plasmid cDNA clones are screened for their ability to select a particular mRNA. Plasmid DNA is bound to nitrocellulose filters, hybridized to mRNA, washed, and the selected mRNA is eluted from the filter. Eluted mRNA is characterized by translation into 35S-labeled protein, which is identified by immunoprecipitation and denaturing (SDS) polyacrylamide gel electrophoresis. The desired clone is that which is able to select an mRNA that translates into the desired protein. This procedure can be modified to characterize cosmids and bacteriophage DNA.