UNIT 11.8 In Vitro Translation Using HeLa Extract
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Cell Biology
How to Cite
Witherell, G. 2001. In Vitro Translation Using HeLa Extract. Current Protocols in Cell Biology. 6:11.8:11.8.1–11.8.10.
- Published Online: 1 MAY 2001
- Published Print: APR 2000
A HeLa cell extract can be used as an alternative to rabbit reticulocyte lysate (RRL) for in vitro translation of mRNAs. It is particularly suited for synthesis of proteins that cannot be translated by RRL, is more cap dependent, has a higher capacity for faithful initiation at the correct start codon, and is more likely representative of translation in intact mammalian cells. This unit provides a detailed protocol for production and use of an mRNA-dependent cell-free translation system from HeLa extract. A support protocol describes production and purification of the mRNA.