In 2009, PacBio published a paper expanding the principle into a full-blown sequencing technique. Once again, each nucleotide was labeled with a different colored fluorophore detectable by the ZMW to determine which base had been incorporated. The fluorophores were attached such that they would be cleaved off during the chemical reaction incorporating the base into the growing DNA strand; they would then diffuse out of the ZMW so that the next fluorophore could be detected. Sequencing took place on a chip with many wells simultaneously — a different type of parallelization where each well detected a single DNA molecule undergoing the same basic chemical reaction.
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Фото: Global Look Press
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