You should see individual, sharp and evenly spaced peaksģ. You can use any of the following programs to view your. Here are a few guidelines to help with DNA sequencing troubleshooting and analysis 1. In fact this is so ambiguous that the DNA sequencing reaction should be repeated. If you never looked at the trace you would be happy.īut look closer, the overlapping peaks in the chromatogram suggest the results are not as certain as the sequence may suggest. Here is an example of a seemingly clean DNA sequence (no Ns in sight). An example of where the chromatogram can come to your rescue for DNA sequencing troubleshooting and analysis
And, like all controls, missing out is a big mistake. When it comes to DNA sequencing the chromatogram is your visual control. These controls help you properly visualize your results. When you run a restriction digest on a gel you always include proper controls like uncut DNA and the proper ladder. The most important of those is to always look closely at the trace file (or chromatogram) of the sequencing results you get back from your favorite sequencing facility.
So I have developed some good habits that I wanted to pass on to you to make sure you are getting the most out of the data you get back from your sequencing runs. As part of my job ensuring plasmid quality at Addgene, I analyze 50-100 sequencing reactions a week.
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