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Cloning and expansion of repetitive DNA sequences
Sophie L Williams1, Gideon Coster1
1Genome Replication lab, Division of Cancer Biology, Institute of Cancer Research, Chester Beatty Laboratories, London, United Kingdom.
Abstract:
Repeat and structure-prone DNA sequences comprise a large proportion of the human genome. The instability of these sequences has been implicated in a range of diseases, including cancers and neurodegenerative disorders. However, the mechanism of pathogenicity is poorly understood. As such, further studies on repetitive DNA are required. Cloning and maintaining repeat-containing substrates is challenging due to their inherent ability to form non-B DNA secondary structures which are refractory to DNA polymerases and prone to undergo rearrangements. Here, we describe an approach to clone and expand tandem-repeat DNA without interruptions, thereby allowing for its manipulation and subsequent investigation.
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