ena-STUDY-CAM-23-01-2020-11:12:53:229-107
Wolbachia genome sequencing
New Wolbachia genomes were obtained from eleven Drosophila species using the protocol described in Ellegaard, Klasson, Näslund, Bourtzis, & Andersson, 2013. Briefly, Wolbachia cells were purified from 20-30 fly embryos that were dechorionated in bleach and homogenized in phosphate-buffered saline. The homogenate was then centrifuged, passed through 5 and 2.7 µm pore size filters and multiple-displacement amplification was performed directly on the bacterial pellet using the Repli-g midi kit (Qiagen) as in Ellegaard et al., 2013. The amplified DNA was finally cleaned using the QIAamp DNA mini kit prior to sequencing. From each DNA sample, 3 kb paired-end and 50 bp paired-end DNA libraries were prepared. These were multiplexed and sequenced on one plate of 454 Roche FLX (University of Cambridge, Department of Biochemistry, UK) and one lane of Illumina HiSeq2000 instruments (The Genome Analysis Centre, Norwich, UK) respectively.Ellegaard KM, Klasson L, Näslund K, Bourtzis K, Andersson SGE. Comparative genomics of Wolbachia and the bacterial species concept. PLoS Genet. 2013;9: e1003381.
New Wolbachia genomes were obtained from eleven Drosophila species using the protocol described in Ellegaard, Klasson, Näslund, Bourtzis, & Andersson, 2013. Briefly, Wolbachia cells were purified from 20-30 fly embryos that were dechorionated in bleach and homogenized in phosphate-buffered saline. The homogenate was then centrifuged, passed through 5 and 2.7 µm pore size filters and multiple-displacement amplification was performed directly on the bacterial pellet using the Repli-g midi kit (Qiagen) as in Ellegaard et al., 2013. The amplified DNA was finally cleaned using the QIAamp DNA mini kit prior to sequencing. From each DNA sample, 3 kb paired-end and 50 bp paired-end DNA libraries were prepared. These were multiplexed and sequenced on one plate of 454 Roche FLX (University of Cambridge, Department of Biochemistry, UK) and one lane of Illumina HiSeq2000 instruments (The Genome Analysis Centre, Norwich, UK) respectively.Ellegaard KM, Klasson L, Näslund K, Bourtzis K, Andersson SGE. Comparative genomics of Wolbachia and the bacterial species concept. PLoS Genet. 2013;9: e1003381.