Author: Cobucci-Ponzano, Beatrice; Conte, Fiorella; Benelli, Dario; Londei, Paola; Flagiello, Angela; Monti, Maria; Pucci, Piero; Rossi, Mosè; Moracci, Marco
Title: The gene of an archaeal a-l-fucosidase is expressed by translational frameshifting Document date: 2006_8_18
ID: 69gftii4_27
Snippet: For real-time PCR experiments total cDNA was obtained using the kit Quantitect RT (Qiagen GmbH, Hilden, Germany) from 500 ng of the same preparation of RNA described above. cDNA was then amplified in a Bio-Rad LightCycler using the DyNAmo HS Syber Green qPCR Kit (Finnzymes Oy, Espoo, Finland). Synthetic oligonucleotides (PRIMM) used for the amplification of a region at the 3 0 of the ORF SSO3060 were as follows: 5 0 -Real: 5 0 -TAAATGGC-GAAGCGATT.....
Document: For real-time PCR experiments total cDNA was obtained using the kit Quantitect RT (Qiagen GmbH, Hilden, Germany) from 500 ng of the same preparation of RNA described above. cDNA was then amplified in a Bio-Rad LightCycler using the DyNAmo HS Syber Green qPCR Kit (Finnzymes Oy, Espoo, Finland). Synthetic oligonucleotides (PRIMM) used for the amplification of a region at the 3 0 of the ORF SSO3060 were as follows: 5 0 -Real: 5 0 -TAAATGGC-GAAGCGATTTTC-3 0 ; 3 0 -Real: 5 0 -ATATGCCTTTGTCGC-GGATA-3 0 for the gene fucA1. 5 0 -GAATGGGGGTGATA-CTGTCG-3 0 and 5 0 -TTTACAGCCGGGACTACAGG-3 0 for the 16S rRNA gene.
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