Purification and mapping of specific mRNAs by hybridization-selection and cell-free translation.
Authors:
- R P RicciardiRRR P Ricciardi
- J S MillerJMJ S Miller
- B E RobertsBRB E Roberts
- 1 October 1979
- journal article
- research article
- Published by National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 76 (10) , 4927-4931
Abstract
We describe a simple procedure for isolating specific mRNAs and for mapping them to the regions of the DNA from which they originate. The method involves hybridization of total cytoplasmic RNA to restriction fragments of DNA that have been fractionated in agarose gels and immobilized on nitrocellulose filters. The hybridization-selected RNAs are eluted and translated in a cell-free system in order to identify their encoded polypeptides. Optimal hybridization and filter washing conditions are given for selection of mRNAs from adenovirus 2-infected cells and transformed cells.Keywords
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