Proceedings of the National Academy of Sciences · 1977 · 8K citations · 11 references
Life SciencesBiologyMolecular BiologyDna RepairNucleic Acid BiochemistryNucleic Acid ChemistryDna ChemistryDna SequencingNucleic Acid LabelingNucleic Acid SequencingNucleic Acid StructureDna Base PairingDna ElectrophoresisDna LabelingDna Sequence AnalysisDna Sequence CompositionNucleic Acid Sequence AnalysisPolyacrylamide Gel ElectrophoresisSequence-specific Dna Cleavage
DNA sequencing can be performed by chemically cleaving a terminally labeled DNA molecule at each base repetition, with fragment lengths indicating base positions. The authors aim to describe reactions that cleave DNA preferentially at guanines, adenines, cytosines and thymines equally, and at cytosines alone. By resolving the products of these four reactions by size using electrophoresis on a polyacrylamide gel, the DNA sequence can be read from the pattern of radioactive bands. The method enables sequencing of at least 100 bases from the labeling point.
DNA can be sequenced by a chemical procedure that breaks a terminally labeled DNA molecule partially at each repetition of a base. The lengths of the labeled fragments then identify the positions of that base. We describe reactions that cleave DNA preferentially at guanines, at adenines, at cytosines and thymines equally, and at cytosines alone. When the products of these four reactions are resolved by size, by electrophoresis on a polyacrylamide gel, the DNA sequence can be read from the pattern of radioactive bands. The technique will permit sequencing of at least 100 bases from the point of labeling.
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