SpringerProtocols: Abstract: Direct DNA Sequencing of PCR Products Using Magnetic Beads

Direct DNA Sequencing of PCR Products Using Magnetic Beads

By: Joakim Lundeberg², Bertil Pettersson², Mathias Uhlén²

Abstract

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The use of magnetic particles in many fields of biochemistry, molecular biology, and medicine has been well documented, and several magnetic particles are now available for diagnostic and cell-separation purposes. The solid-phase approaches have improved robustness by their increased reproducibility with accompanying higher yields. Furthermore, automation has been facilitated since reaction buffers and additional reagents can be rapidly changed without centrifugation or precipitation steps. The introduction of magnetic particles has proven to be an attractive alternative to traditional means of performing DNA sequencing.

Affiliation(s): (2) Department of Biochemistry and Biotechnology, Royal Institute of Technology, Stockholm, Sweden

Book Title: PCR Sequencing Protocols

Series: Methods in Molecular Biology | Volume: 65 | Pub. Date: Aug-19-1996 | Page Range: 57-66 | DOI: 10.1385/0-89603-344-9:57

Subject: Cell Biology

References

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1.	Sanger, F., Nicklen, S., and Coulson, A. R. (1977) DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci. USA 74(12), 5463–5467.

2.	Scharf, S. J., Horn, G. T., and Erlich, H. A. (1986) Direct cloning and sequence analysis of enzymatically amplified genomic sequences. Science 233, 1076–1078.

3.	Hultman, T., Ståhl, S., Hornes, E., and Uhlén, M. (1989) Direct solid phase sequencing of genomic and plasmid DNA using magnetic beads as solid support. Nucleic Acids Res. 17, 4937–4946.

4.	Hultman, T., Bergh, S., Moks, T., and Uhlén, M. (1991) Bidirectional solid-phase sequencing of in vitro-amplified plasmid DNA. BioTechniques 10(1), 84–93.

5.	Innis, M. A., Myambo, K. B., Gelfand, D. H., and Brow, M. A. D. (1988) DNA sequencing with Thermus aquaticus DNA polymerase and direct sequencing of PCR-amplified DNA. Proc. Natl. Acad. Sci. USA 85, 9436–9440.

6.	Leren, T. P., Rødningen, O. K., Røsby, O., Solberg, K., and Berg, K. (1993) Screening for point mutations by semi-automated DNA sequencing using Sequenase and magnetic beads. BioTechniques 14(4), 618–623.

7.	Khurshid, F. and Beck, S. (1993) Error analysis in manual and automated DNA sequencing. Anal. Biochem. 208, 138–143.

8.	Carothers, A. M., Urlaub, G., Mucha, J., Grunberger, D., and Chasin, L. A. (1989) Point mutation analysis in mammalian gene: Rapid preparation of total RNA, PCR amplification of cDNA, and Taq sequencing by a novel method. BioTechniques 7, 494–499.

9.	Hedrum, A., Pontén, F., Ren, Z., Lundeberg, J., Pontén, J., and Uhlén, M. (1994) Sequence-based analysis of the human p53 gene based on microdissection of tumor biopsy samples. BioTechniques 17, 1–9.

10.	Wahlberg, J., Albert, J., Lundeberg, J., Cox, S., Wahren, B., and Uhlen, M. (1992) Dynamic changes in HIV-1 quasispecies from azidothymidine (AZT) treated patients. FASEB J. 6, 2843–2847.

11.	Leitner, T., Halapi, E., Scarletti, G., Rossi, P., Albert, J., Fenyo, E.-M., and Uhlén, M. (1993) Analysis of heterogeneous viral populations by direct DNA sequencing. BioTechniques 15(1), 120–427.

12.	Larder, B. A., Kohli, A., Kellam, P., Kemp, S. D., Kronick, M., and Henfrey, R. D. (1993) Quantitative detection of HIV-1 drug resistance mutations by automated DNA sequencing. Nature 365, 671–673.

13.	Holmberg, A., Fry, G., and Uhlén, M. (1993) Automated preparation of DNA templates for sequencing on the ABI Catalyst robotic workstation, in Automated DNA Sequencing and Analysis Techniques (Venter, C., ed.), Academic, London, pp. 139–145.

14.	Wahlberg, J., Albert, J., Lundeberg, J., Fenyo, E.-M., and Uhlén, M. (1991) Analysis of the V3 loop in neutralization-resistant human immunodeficiency virus type 2 variants by direct solid phase DNA sequencing. AIDS Res. Hum. Retrov. 7(12), 983–990.

15.	Wahlberg, J., Lundeberg, J., Hultman, T., and Uhlén, M. (1990) General colorimetric method for DNA diagnostics allowing direct solid phase genomic sequencing of the positive samples. Proc. Natl. Acad. Sci. USA 87, 6569–6573.

16.	Pettersson, B., Johansson, K.-E., and Uhlén, M. (1994) Sequence analysis of 16S rRNA from Mycoplasmas by direct solid-phase DNA sequencing. Appl. Env. Microbiol. 60(7), 2456–2461.

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