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Jenna M. Lang edited Methods.md
over 9 years ago
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Genomic DNA was extracted using a Wizard Genomic DNA Purification Kit (Promega). A nearly full-length 16S sequence was amplified using the 27F (5'-AGAGTTTGATCMTGGCTCAG-3') and 1391R (5'-GACGGGCGGTGTGTRCA-3') "universal" primers. Sanger sequencing was provided by the College of Biological Science UC-DNA Sequencing Facility (UC Davis). This DNA was also used for Illumina sequencing of the draft genome as described elsewhere (Coil and Eisen, STATUS?). The genome sequence was annotated using the RAST server \cite{Aziz_2008} \cite{Overbeek_2013}.
A phylogentic tree was
constructed inferred using all type strains from the _Erythrobacteraceae_ family. The 1491bp 16S rDNA sequence was obtained from the genome assembly in RAST, and uploaded to the Ribosomal Database Project
(RDP) \cite{Cole_2013}.
An alignment RDP was
created used to build an alignment including every Type Strain within the _Erythrobacteraceae_ family,
including strain Coronado(T). Because the taxon names exported with this alignment contained special characters that were not compatible with phylogenetic reconstruction software, a custom script was used to remove or replace those characters with underscores (PUT SCRIPT ON GITHUB AND WITH SOME DOCUMENTATION ABOUT HOW TO RUN IT AND INCLUDE LINK). FastTree \cite{19377059} was used with default settings to build an unrooted phylogeny. Dendroscope 3 \cite{22780991} was used to view and edit the phylogenetic tree in order to 1) root the tree with a clade that is sister to a well-supported clade containing strain Coronado(T),
2) prune the tree to remove taxa outside the clade containg the outgroup, and
??? as an outgroup. METHODS FOR TREE FROM HERE. USED JENNA'S SCRIPT, THEN FASTTREE, THEN DENDROSCOPE. 3) "prettify" the tree.