Re:Bottleneck is somewhere else... (Score 1)

While I agree that the bottleneck is elsewhere, I do not think they use the procedure you mention. If some of them do cycle sequencing, remember that it is not exponential (unlike PCR). There is no reason to use a thermostable polymerase for anything else here, and I would bet they do 'old-fashioned' sequencing with T7. And if they use restriction enzymes I would bet on SauIII because it might be used to generate more or less random fragments. Other might be useless, because they are not working with plasmids! The sequencing method you mention seem sth like Maxam-Gilbert, but you do not seem to remember it well. You do not need external standards (nor internal) for DNA sequencing, and they certainly use the Sanger dideoxy method (Maxam is usually used in footprinting experiments). Finally nobody does radiographies on film now. You just fluorescence detection in an automatic sequencing aparatus. I definitely agree with the conclusion, it takes more human-power than computer-power to complete the project.