While Sanger sequencing was highly accurate and less labor-intensive than its predecessors, it still required the use of radioactive reagents and manual sequence recording. In 1986, Leroy Hood’s lab at Caltech replaced the radiolabeled ddNTPs with fluorescently labeled nucleotides, using fluorophores that emitted different colors of light for each base. They were now able to run the products of all four reactions on the same gel and have a computer read the sequence by detecting the color of each fluorescent signal as fragments passed through a laser beam.
It was a severe limitation.
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