Genome sequence of Rhodopseudomonas palustris: environmental isolate: Sample Rhodopseudomonas palustris WS17
"Genomic DNA was sheared using a Bioruptor UCD-200. Following shearing, samples were purified and fragment size was assessed by PAGE Gel Electrophoresis. Sheared samples were repaired using the Epicentre End-It DNA End Repair kit. Following End Repair, samples were re-purified. Each library was subjected to A-Tailing. Adaptor oligonucleotides were used in a ""Y"" conformation, achieved by annealing of oligonucleotides containing partial complimentary sequences, in combination with a 5' phosphate and a 3' phosphoorthiolation of the two oligonucleotides, to ensure each A-tailed fragment strand receives the forward and reverse priming sequences on opposite ends.  Adaptor oligonucleotides were ligated in a molar ratio of 10 moles of the appropriate barcoded ""Y"" adaptor to 1 mole of A-tailed fragment ends. Ligation results were subjected to agarose-gel size selection. Products in the desired size range (which varies depending on the experimental goal) were excised using disposable Safe Xtractors tools. Size selected fragments were amplified using stock amplification primers. Amplified products were purified using the Qiagen Min-Elute PCR purification kit. Products were quantified using the Quantifluor ST (see above) and/or the Agilent Bioanalyzer. Average fragment size was visually confirmed using PAGE analysis, and/or the Agilent Bioanalyzer."