Interviewee: Michael Hunkpiller.
Michael Hunkapiller talks about the process of developing the automated sequencing machine.
(DNAi Location: Manipulation > Techniques > Sorting & sequencing > Making sequencing automated)
The method of doing DNA sequencing at all in a routine fashion was pretty new at that point. The original papers were published in '77. And it was pretty clear that if you wanted to tackle problems in a robust way, and with a lot of scale, that there were some inherent problems with the technique. The chemistry was a nice elegant approach, it really solved a lot of clever problems that way that weren't solvable in the protein space. But it was a tool that used radioactivity as its detection methodology, and there are limits as to how much you want to scale that up, just from the hazardous requirements that are involved in handling it. It was a very person-intensive process; the whole procedure was, was manual by its very nature and, worse, the interpretation of the data was very subjective. And what we thought we needed to do was come up with an analytical technique, playing off the Sanger chemistry, that allowed a computer to be able to read and interpret the data, because that's the only way you're ever going to scale up the automation of a big process.
The sequencing method developed by Fred Sanger forms the basis of automated "cycle" sequencing reactions today. Fluorescent dyes are added to the reactions, and a laser within an automated DNA sequencing machine is used to analyze the DNA fragments produc
Two sequencing techniques were developed independently in the 1970s. The method developed by Fred Sanger used chemically altered "dideoxy" bases to terminate newly synthesized DNA fragments at specific bases (either A, C, T, or G). These fragments are th