Partial-digest DNA sequencing.
A technique called partial-digest sequencing that permits DNA of 4-6 kb in length to be sequenced without subcloning is described. The method exploits the specific cuts introduced by partial digestion with restriction endonucleases that have 4-base recognition sites to produce ordered ladders of PCR-amplified fragments. The staggered ends contain PCR primers and can thus be individually sequenced using conventional methods to yield overlapping sequences covering the entire region. This method should have significant impact on both large and small DNA sequencing projects and find many applications in general manipulations in which ordered sets of deletions need to be produced.
Wright
W E
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Department of Cell Biology and Neuroscience, University of Texas Southwestern Medical Center, Dallas 75235.
D000818:Animals; D001483:Base Sequence; D004277:DNA, Single-Stranded; D008969:Molecular Sequence Data; D009124:Muscle Proteins / Q000235:genetics; D018008:Myogenin; D010957:Plasmids; D016133:Polymerase Chain Reaction; D051381:Rats; D017422:Sequence Analysis, DNA / Q000379:methods*
D004277:DNA, Single-Stranded; D009124:Muscle Proteins; C503803:Myog protein, rat; D018008:Myogenin