Abstract
Ligase chain reaction (LCR), employing just oligonucleotide probes and Principle and DNA ligase, is capable of detecting approximately <_ 1000 copies of a specific applications target DNA sequence in the presence of a vast excess of other DNA sequence information. Since the first description in 1989 (Backman and Wang, 1989; Royer et al., 1989; Wallace, 1989; Wu and Wallace, 1989; Orgel, 1989; Richards and Jones, 1989) LCR has been improved by the employment of a thermostable DNA ligase in conjunction with nonradioactive detection (Bond et al., 1990). The ability of LCR to distinguish between normal βAand sickel βS-globin genotypes from 10 µl of whole blood using probes having a mismatch or full complementarity at the point of ligation has recently been reported (Barany, 1991), demonstrating the potential of LCR in allele-specific detection.
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References
Backman KC, Wang C-NJ (1989) Method for detecting a target nucleic acid sequence. Eur Pat Appl 0 320 308
Barany F (1991) Genetic disease detection and DNA amplification using cloned thermostable ligase. Proc Natl Acad Sci USA 88:189–193
Barker DG, Johnson AL, Johnson LH (1985) An improved assay for DNA ligase reveals temperature-sensitive activity in cdc9 mutants ofSaccharomyces cerevisiae.Mol Gen Genet 200:458–462
Nickerson DA, Kaiser R, Lappin S, Stewart J, Hood L, Landegren U (1990) Automated DNA diagnostics using an ELISA-based oligonucleotide ligation assay. Proc Natl Acad Sci USA 87:8923–8927
Orgel LE (1989) Ligase based amplification method. PCT Int Appl WO 89/09835
Richards RM, Jones T (1989) Method and reagents for detecting nucleic acid sequences. PCT Int Appl WO 89/12696
Royer GP, Cruickshank KA, Morrison LE (1989) Template-directed photoligation. Eur Pat Appl 0 324 616
Shimer Jr. GH, Backman KC (1995) Ligase chain reaction. In: Howard JJ, Walker JM (eds) Methods in Molecular Biology, Vol. 46. Humana Press, Boca Raton, FL, pp 269–278
Wallace BR (1989) Method of amplifying and detecting nucleic acid sequences. Eur Pat Appl 0 336–731
Wu DY, Wallace RB (1989) The ligation amplification reaction (LAR): Amplification of specific DNA sequences using sequential rounds of template-dependent ligation. Genomics 4:560–569
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© 2000 Springer-Verlag Berlin Heidelberg
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Jr Shimer, G.H., Backman, K.C. (2000). Ligase Chain Reaction. In: Kessler, C. (eds) Nonradioactive Analysis of Biomolecules. Springer Lab Manuals. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-57206-7_32
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DOI: https://doi.org/10.1007/978-3-642-57206-7_32
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-540-64601-3
Online ISBN: 978-3-642-57206-7
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