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Using SD-AFLP and MSAP to assess CCGG methylation in the banana genome
Authors:Franc-Christophe Baurens  François Bonnot  David Bienvenu  Sandrine Causse  Thierry Legavre
Institution:(1) CIRAD AMIS Biotrop, UMR 1098 (BEPC), TA 40-03, Avenue Agropolis, 34398 Montpellier Cedex 5, France;(2) CIRAD CP, TA 80-03, Avenue Agropolis, 34398 Montpellier Cedex 5, France;(3) CIRAD FLHOR, UMR 1098 (BEPC), TA 50/PS4 Avenue Agropolis, 34398, Montpellier Cedex 5, France;(4) CIRAD AMIS Biotrop, UMR 1098 (BEPC), TA 40-03, Avenue Agropolis, 34398 Montpellier Cedex 5, France
Abstract:Two amplified fragment length polymorphism (AFLP)-derived techniques were used to assess methylation at CCGG sites in the banana genome. Assessment of these techniques revealed that, while amplification steps are very reproducible, the ligation step is more subject to variability. Overall, these techniques produced an error rate of 0.2% per analysed band. Statistical approach highlights the fact that sample duplication is necessary to produce reliable results. This study involved 18 primer pairs and found that in the banana genome, roughly 80% of CCGG sites are unmethylated, 5% are methylated at the internal cytosine, and 15% are methylated at the external or both cytosines.
Keywords:amplified fragment length polymorphism  banana  DNA methylation  methylation-sensitive amplification polymorphism  secondary digest-AFLP
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