Translational extracts active biologically in vitro obtained from eukaryotic monolayer cells: a versatile method for viral RNA studies

Preservation of enzymatic activities in biological samples, especially afte r freeze/thawing, is a crucial requirement in virological research. Theoret ically, this preservation can be achieved with the presence of cryopreserva tive agents. In contrast to tedious methods, it was found that this might b e readily achieved by using well-defined conditions, including sucrose in t he samples. Hence, the generation of a translational extract obtained from eukaryotic cells that have grown as monolayers is described below. This ver satile method could be used advantageously for the in vitro translation of messenger RNAs, added exogenously, including viral mRNAs. The translational extract can be prepared freshly on a daily basis, or more conveniently it can be frozen and thawed subsequently for further use, without loss of acti vity. It can replace the Krebs ascites fluid and the commercial rabbit reti culocyte lysate. The procedure employed for the preservation of the biologi cal activity of the translational extract can be extended to various other biological samples. (C) 2001 Elsevier Science B.V. All rights reserved.