Abstract
In 1993 the etiology of Huntington’s disease (HD) was revealed as an expansion of the trinucleotide sequence CAG on chromosome 4p16.3. Normal individuals have between 10 and 29 repeats. An intermediate range has been established between 30 and 35 CAG repeats. Clinical manifestation of the disease occurs with 36–121 repeats, the median being about 44 [Huntington’s Disease Collaborative Reasearch Group, 1993]. Since the causal discovery of the CAG repeat, PCR amplification and subsequent post-PCR gel sizing of the product has been the gold standard for the diagnosis of HD [Andrew, 1994]. We present a rapid PCR protocol wherein the Tm of the PCR product of the CAG repeat region can be used for Huntington’s diagnosis. Melting curve analysis in the presence of SYBR Green I is used to determine the product Tm that is dependent on product length. In addition, we compared melting curves obtained on the ABI 5700 with those obtained on the LightCycler instrument.
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© 2002 Springer-Verlag Berlin Heidelberg
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Gundry, C.N., Wittwer, C.T. (2002). SYBR Green I Analysis of the Trinucleotide Repeat Responsible for Huntington’s Disease. In: Dietmaier, W., Wittwer, C., Sivasubramanian, N. (eds) Rapid Cycle Real-Time PCR — Methods and Applications. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-59397-0_7
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DOI: https://doi.org/10.1007/978-3-642-59397-0_7
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-642-63965-4
Online ISBN: 978-3-642-59397-0
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