![]() The distribution of Ilarvirus species populations amongst 61 Australian Prunus trees was determined by next generation sequencing (NGS) of amplicons generated using a genus-based generic RT- PCR targeting a conserved region of the Ilarvirus RNA2 component that encodes the RNA dependent RNA polymerase (RdRp) gene. Kinoti, Wycliff M Constable, Fiona E Nancarrow, Narelle Plummer, Kim M Rodoni, Brendan ![]() Generic Amplicon Deep Sequencing to Determine Ilarvirus Species Diversity in Australian Prunus. ![]() In this project, deep amplicon sequencing was used to generate 50,926 phytoplasma sequences from 11 phytoplasma-infected grapevine samples from a PCR amplicon in the 5' end of the 16S region. The diversity of phytoplasmas within single plants has not yet been fully investigated. Nicolaisen, Mogens Contaldo, Nicoletta Makarova, Olga Thus, single strand dideoxynucleotide sequencing with adenylated primers enables complete or near complete readability of short PCR amplicons.ĭeep amplicon sequencing reveals mixed phytoplasma infection within single grapevine plants In addition, in the case of TGFB1 products (366 bp, 432 bp, and 680 bp, respectively, the lengths of sequencing readings were significantly longer if adenylated primers were used. Complete sequences of shorter products (AMELX 106 bp, SERPINA1 121 bp, HFE 208 bp, APOE 244 bp, MBL2 317 bp were obtained. Polyadenylation of the sequencing primers minimized the loss of bases in all amplicons. ![]() Performing a polymerase chain reaction, we amplified eight amplicons of six human genes (AMELX, APOE, HFE, MBL2, SERPINA1 and TGFB1 ranging from 106 bp to 680 bp. We present a method which extends the readability by using sequencing primers modified by polyadenylated tails attached to their 5' ends. Loss of an initial part of nucleotides behind the 3' end of the sequencing primer limits the readability of sequenced amplicons. Polyadenylated Sequencing Primers Enable Complete Readability of PCR Amplicons Analyzed by Dideoxynucleotide Sequencingĭirectory of Open Access Journals (Sweden)įull Text Available Dideoxynucleotide DNA sequencing is one of the principal procedures in molecular biology. ![]()
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |