caa a22 4500 465795080 CHVBK 20180323112035.0 cr unu---uuuuu 170327e19900601xx s 000 0 eng 10.1007/BF00308566 doi (NATIONALLICENCE)springer-10.1007/BF00308566 A possible approach to enrich cDNA yields with full-length molecules [Elektronische Daten] [Vladimir Zaslavsky, Theah Molad] Newcastle disease virus (NDV)-specific hemagglutinin-neuraminidase (HN) mRNAs were used as the model templates for cDNA synthesis. Polyadenylated RNAs were isolated from the particulate fraction of cytoplasmic extracts of NDV-infected cells rather than from nonfractionated extracts. The approach is based on earlier findings that eucaryotic mRNAs are present in cytoplasmic extracts in the form of ribonucleoproteins (mRNPs) rather than as free nucleic acids. The idea of the approach was to separate mRNPs from cell sap RNases prior to RNA extraction in order to minimize partial enzymatic hydrolysis of mRNAs. The presence of the 5′ terminus (mRNA sense) in cDNAs synthesized was considered as an indication for the suitability of mRNA templates for cDNA synthesis. The cDNAs were synthesized and cloned in lambda gt10 phage. About 300 phages carrying the HN-specific inserts have been identified among 50,000 recombinants, and nine of them were analyzed for the presence of the HN 5′ terminus. It was found that the termini are present in all the clones analyzed. The result is in an agreement with the expectation that removal of cell sap prior to RNA extraction significantly increases the suitability of RNA templates for cDNA synthesis. Kluwer Academic Publishers, 1990 cDNA synthesis nationallicence mRNA nationallicence NDV nationallicence hemagglutinin-neuraminidase gene nationallicence Zaslavsky Vladimir Kimron Veterinary Institute, Bet Dagan, Israel aut Molad Theah Kimron Veterinary Institute, Bet Dagan, Israel aut Virus Genes Kluwer Academic Publishers 4/1(1990-06-01), 63-72 0920-8569 4:1<63 1990 4 11262 https://doi.org/10.1007/BF00308566 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00308566 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Zaslavsky Vladimir Kimron Veterinary Institute, Bet Dagan, Israel aut NATIONALLICENCE 700 1- Molad Theah Kimron Veterinary Institute, Bet Dagan, Israel aut NATIONALLICENCE 773 0- Virus Genes Kluwer Academic Publishers 4/1(1990-06-01), 63-72 0920-8569 4:1<63 1990 4 11262 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer