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Strategy for Construction of a cDNA Encoding a Repetitive Amino Acid Sequence

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Generation of cDNA Libraries

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 221))

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Abstract

The polymerase chain reaction (PCR) is a powerful method permitting generation of almost any desired cDNA sequence. Construction of cDNA fragments that encode repetitive amino acid sequences by PCR has proven problematic, however, as repetitive nucleotide primers tend to amplify undesired fragments (because of their misannealing with the template). To overcome this problem, Dombrowski and Wright (1) developed a solid-phase gene-assembly protocol, but because this technique requires preparation of a solid-phase reaction system, it cannot be routinely performed in most laboratories. Here, we describe a new strategy that makes use of standard PCR protocols, yet enables assembly of cDNAs encoding repetitive amino acid sequences (2).

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References

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© 2003 Humana Press Inc., Totowa, NJ

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Asada, M., Imamura, T. (2003). Strategy for Construction of a cDNA Encoding a Repetitive Amino Acid Sequence. In: Ying, SY. (eds) Generation of cDNA Libraries. Methods in Molecular Biology™, vol 221. Humana Press. https://doi.org/10.1385/1-59259-359-3:261

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  • DOI: https://doi.org/10.1385/1-59259-359-3:261

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-066-3

  • Online ISBN: 978-1-59259-359-0

  • eBook Packages: Springer Protocols

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