Figure 1 : Preparation and sequencing of AFSM tags.

From: AFSM sequencing approach: a simple and rapid method for genome-wide SNP and methylation site discovery and genetic mapping

Figure 1

Sample preparation for AFSM genotyping is accomplished by combining two restriction enzyme pairs (EcoRI-MspI and EcoRI-HpaII) to digest genomic DNA and incorporating barcodes for multiplex sequencing. EcoRI is used as a rare cutter, and the methylation-retraction enzymes HpaII and MspI are employed as frequent cutters. HpaII and MspI have different sensitivities to methylation of the inner or outer cytosines and can produce different products, reflecting the different methylation states of the cytosines.