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A versatile primer for DNA sequencing in the M13mp2 cloning system |
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| Authors: | G Heidecker J Messing B Gronenborn |
| Institution: | 1. Department of Bacteriology, University of California, Davis CA 95616 U.S.A.;1. Institut für Genetik, Universität Köln, Weyertal 121, D 5000 KölnF.R.G. |
| Abstract: | A primer for DNA sequencing by the chain-termination method in the M13mp2 cloning system was constructed and amplified. The primer was isolated as an EcoRI/AluI restriction fragment. After conversion of the AluI end into an EcoRI end the fragment was cloned in pBR325 from which it can be recovered by cleavage with EcoRI. The primer hybridizes to the single-stranded DNA of the mature M13mp2 phage next to the site of insertion thereby directing DNA synthesis along the inserted DNA. |
| Keywords: | Single-stranded DNA phage dideoxy chain terminators conversion of restriction sites Ap ampicillin bp base pairs Cm chloramphenicol dd dideoxy DTT dithiothreitol Tc tetracycline |
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