Methyl-CpG-Binding PCR of Bloodspots for Confirmation of Fragile X Syndrome in Males
Factors critical for the detection specificity of MB-PCR. (a) NaCl concentration in the stringency wash and enzymatic cleavage of the test DNA are the major factors that can critically determine the capability of MB-PCR in distinguishing between samples with methylated and unmethylated FMR1 alleles. (b) Under the same MB-PCR condition, including a stringency wash with 400 mM NaCl and 45 PCR cycles of amplification, the samples cleaved by different restriction enzyme(s) varied considerably in the detection sensitivity. (–) Control DNA with universally unmethylated sequences; (+): control DNA with universally methylated sequences; (M): molecular-size markers
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