53 Analyzing Chromatograms to Confirm Consensus Sequence

Chromatograms from Sanger DNA sequencing can give us helpful information about how to assemble a consensus sequence from multiple different sequencing reads along a gene of interest. Recall we performed sequencing reactions using primers located at positions 27 (forward primer), 515 (forward primer), and 1492 (reverse primer). There will be overlapping sequences between primers 27 and 515, as well as overlapping sequences from 515 and 1492. You can use these to disregard poor-quality base calls that occur at the beginning and end of Sanger sequencing reactions to assemble a faithful, high-quality sequence to BLAST to identify your bacterial species preliminarily. The variable regions of the 16S gene differ between species and can help us identify our bacterial isolates. We will focus on those to identify the genus and species of the isolates you have been working with!