DNA Sequencing and Extraction of Deinococcus caeni
Introduction: Nanopore sequencing with Deinococcus caeni has been the goal for the past week or so in the lab, but this week it came to fruition unsuccessfully. The DNA extracted last week was processed in the Nanopore Ligation V14 kit and sequenced attempted, and then post sequencing sample analysis of approximate genome size for more in depth analysis in preparation for the next sequencing attempt. Methods: The first step in preparing for sequencing was to do a DNA cleanup and concentration of the sample produced from the previous week. The NEB Monarch DNA cleanup kit was utilized by diluting the 18 ul of sample to a total volume of 50 ul with a 1:2 binding buffer ratio to select for genomic DNA at higher kbp fragments. The protocol was followed until the elution step in which 15 ul of PCR water was used as the eluent and an incubation step at room temperature for five minutes prior to centrifugation. The sample was tested for concentration and cleanliness on the Nanopore ...