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Observed solution Tm of partially matched probe-target duplexes with varying complementary stretch. Complementary stretches were positioned at central (indicated by blue) or terminal ends (indicated by red). Stretch length varied from 9 to 21nt. Each target was mixed with the probe to form a duplex and then allowed to undergo melting followed by denaturation to obtain melting temperature. Error bars are standard deviation among replicates for probe-target duplex.
