Horizon 1 1 17 ACL : Articles dans des revues avec comité de lecture répertoriées par l'AERES Biochemical and Biophysical Research Communications 1080-1085 sequencing PCR RNA viruses LoPPS 2006 fdi:010037790 ENG Biochemical and Biophysical Research Communications 0006-291X CC:0002376792-0004 4 10.1016/j.bbrc.2006.04.015 https://www.documentation.ird.fr/hor/fdi:010037790 https://www.documentation.ird.fr/intranet/publi/2007/suppl/010037790.pdf 344 Horizon (IRD) Here, we propose an optimised protocol (LoPPS, long PCR product sequencing) which allows the fast, cost-attractive, and high-throughput sequencing of long PCR products. LoPPS constitutes an alternative to the primer-walking technology which is expensive and time consuming but remains the current standard procedure. It is based on the ultrasonic shearing, polishing, and cloning of PCR or RT-PCR products and is compatible with 96- or 384-well microplate systems in which bacterial growth, preparation of plasmid DNA, and sequencing can be automated. We present results obtained from 24 different RT-PCR products (2.5-4.8 kbp long) obtained from various RNA viruses and fully sequenced using LoPPS. The method proved to be robust and fast. It was successfully used on a low amount of DNA and allowed each target nucleotide position to be controlled twice or more, with a final cost which is one-third of that of primer-walking. 052 ; 020