The QIAGEN Guide to Template Purification and DNA Sequencing

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Sequences of 1000-bp PCR fragment contaminated with the amplification primer and sequenced with a nested primer. A contamination with 1 pmol of each amplification primer. Note the reduced sequence quality. B contamination with 2 pmol of each amplification primer. This gives an unreadable sequence. ᭿ ᭿ 36 VIII. 5 and 100 µM of each dNTP (Figure 28). Contamination with nucleotides at concentrations of 10 µM or more led to a decrease in signal strength and a greater number of ambiguities. An increase in the proportion of long fragments was also observed.

Academic Press, New York, pp 211–215. 83. M. (1996) BioTechniques 20, 862. 84. A. et al, PE-Applied Biosystems (1998) Poster presentation, Microbial Genomes II Meeting, Hilton Head, USA, 31 January – 3 February. 85. , Plunkett, G. , Shao, Y. (1997) Science 277, 1453. © 1998 QIAGEN XI. qxd 3/6/98 17:06 Uhr Page 50 XII. Appendix: Protocols and Technical Information A. Sequencing kits and reagents C. Cycle sequencing parameters Kits were used according to the suppliers’ (Applied Biosystems) recommendations.

VIII. qxd 3/6/98 17:06 Uhr Page 46 IX. Conclusions Since the publication of the first edition of The QIAGEN Guide to Template Purification and DNA Sequencing, changes in sequencing chemistry and instrumentation have significantly improved both the power, sensitivity, and reliability of automated DNA sequencing. At the same time, the pace of research utilizing DNA sequence data has accelerated, leading to ever higher demands for increased throughput and convenience. QIAGEN will continue to support researchers using DNA sequencing by providing efficient, costeffective methods for purifying template DNA and automating laboratory procedures.

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