Manual editing of the sequence data does improve, sometimes dramatically, the accuracy of a sequence. Most of the data returned used ABI instruments and software, and is reflective of that system's strengths and weaknesses. This test sequence included sequence stretches that are extremely difficult for the TaqFS dye terminator system to read correctly. However, the electropherogram patterns are reproducible and easily learned through experience, and the improvement in sequence accuracy due to editing clearly shows this. The number of labs that submitted both edited and non-edited data is large enough that we should be able to make a statistically significant statement on editing.
Conclusions regarding the effects of altered reaction conditions are very preliminary.
When the data from all the laboratories are pooled, it seems apparent that DMSO can improve, sometimes significantly, the accuracy of the basecalling. This trend holds for all sequence intervals examined. Assessing the effectiveness of altered thermocycling is somewhat unclear due to the fact that different labs use different protocols. Further analysis of altered thermocycling will be conducted and incorporated into this study. Looking at the rankings of the best sequences and the conditions used to generate them, there are a variety of conditions which can produce very good sequence.