Abstracts
Being very useful and informative, many techniques based on nucleic acids hybridization suffer from the cross-annealing of repetitive DNA, presenting in reassociating samples. This “wrong” annealing causes “nonspecific” hybridization of nonorthologous DNA fragments, thus producing chimeric sequences and at the final stage significantly hampering the analysis of the resulting cDNA or genomic libraries. Such chimeras may constitute up to 40–60% of DNA libraries. Importantly, the number of chimerical clones positively correlates with the complexity of hybridizing genomic or cDNA mixtures. The hybridization specificity is a crucial factor determining both the fidelity the efficiency of all hybridizationbased analytical techniques. In this chapter, I review the current attempts to increase the specificity of hybridization at both stages: during nucleic acids reassociation and at the stage of selection of proper hybrids. To this end, approaches based on chemical modifications, improving hybridization kinetics, and improving selection of perfectly matched duplexes, have been developed.
Keywords
- Hybridization specificity
- PCR selection effect
- targeted genomic difference analysis (TGDA)
- thermodynamically stable duplexes
- phenol emulsion reassociation technique (PERT)
- repetitive element
- genomic repeat
- chimerical clone
- hybridization temperature
- melting point
- perfectly matched hybrids
- mispaired DNA rejection (MDR)
- TILLING
- mismatch-sensitive nuclease
- mung bean nuclease
- CotA fraction
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© 2007 Springer
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Buzdin, A.A. (2007). Current Attempts to Improve the Specificity of Nucleic Acids Hybridization. In: Buzdin, A.A., Lukyanov, S.A. (eds) Nucleic Acids Hybridization Modern Applications. Springer, Dordrecht. https://doi.org/10.1007/978-1-4020-6040-3_10
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DOI: https://doi.org/10.1007/978-1-4020-6040-3_10
Publisher Name: Springer, Dordrecht
Print ISBN: 978-1-4020-6039-7
Online ISBN: 978-1-4020-6040-3
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