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Clinical Chemistry, Vol 35, 2196-2201, Copyright © 1989 by American Association for Clinical Chemistry
LJ McBride, SM Koepf, RA Gibbs, W Salser, PE Mayrand, MW Hunkapiller and MN Kronick
Applied Biosystems, Foster City, CA 94404.
Polymerase chain reaction (PCR) as a method for preparing DNA templates has been used for several DNA sequencing applications. An in situ procedure for directly sequencing PCR products by the dideoxy- termination method has been developed by using fluorophore-labeled sequencing primers. Completed sequence reactions were combined and loaded into a single electrophoretic lane of a fluorescence-based DNA sequence analyzer. DNA targets devoid of a universal primer sequence could be sequenced with labeled universal primers by incorporating a universal primer sequence into the PCR product. With this method, the sequence of a 351-bp region in the bacteriophage lambda genome was fully analyzed in a single lane with automatic base identification accuracy of greater than 99%. An unknown sequence, 1.7 kb long, also was sequenced by this procedure, in combination with a "PCR gene walking" strategy. Comparison of the 1110 bases in overlapping sequence data from both strands yielded only two single-base ambiguities. Automated DNA sequence analysis of the highly polymorphic HLA-DQA-1 (alpha) region in the human genome can be performed with this simple methodology. Use of this PCR-sequencing method to analyze DNA extracted from a one-month-old blood sample from an individual who is heterozygous at this locus allowed unambiguous assignment of genotype.
The following articles in journals at HighWire Press have cited this article:
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  B. P. Lomans, P. Leijdekkers, J.-J. Wesselink, P. Bakkes, A. Pol, C. van der Drift, and H. J. M. O. den Camp Obligate Sulfide-Dependent Degradation of Methoxylated Aromatic Compounds and Formation of Methanethiol and Dimethyl Sulfide by a Freshwater Sediment Isolate, Parasporobacterium paucivorans gen. nov., sp. nov. Appl. Envir. Microbiol., September 1, 2001; 67(9): 4017 - 4023. [Abstract] [Full Text] [PDF]
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B. P. Lomans, R. Luderer, P. Steenbakkers, A. Pol, C. van der Drift, G. D. Vogels, and H. J. M. Op den Camp Microbial Populations Involved in Cycling of Dimethyl Sulfide and Methanethiol in Freshwater Sediments Appl. Envir. Microbiol., March 1, 2001; 67(3): 1044 - 1051. [Abstract] [Full Text]
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 P. Crisanti, B. Omri, E. J. Hughes, G. Meduri, C. Hery, E. Clauser, C. Jacquemin, and B. Saunier The Expression of Thyrotropin Receptor in the Brain Endocrinology, February 1, 2001; 142(2): 812 - 822. [Abstract] [Full Text] [PDF]
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 F. H. Crocker, J. K. Fredrickson, D. C. White, D. B. Ringelberg, and D. L. Balkwill Phylogenetic and physiological diversity of Arthrobacter strains isolated from unconsolidated subsurface sediments Microbiology, June 1, 2000; 146(6): 1295 - 1310. [Abstract] [Full Text]
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B. P. Lomans, R. Maas, R. Luderer, H. J. M. Op den Camp, A. Pol, C. van der Drift, and G. D. Vogels Isolation and Characterization of Methanomethylovorans hollandica gen. nov., sp. nov., Isolated from Freshwater Sediment, a Methylotrophic Methanogen Able To Grow on Dimethyl Sulfide and Methanethiol Appl. Envir. Microbiol., August 1, 1999; 65(8): 3641 - 3650. [Abstract] [Full Text]
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 P Fortina, G Dotti, R Conant, G Monokian, T Parrella, W Hitchcock, E Rappaport, E Schwartz, and S Surrey Detection of the most common mutations causing beta-thalassemia in Mediterraneans using a multiplex amplification refractory mutation system (MARMS). Genome Res., November 1, 1992; 2(2): 163 - 166. [Abstract] [PDF]
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