I used Qiagen miRNEasy micro kit, miScript RT II kit and miScript SYBR greenPCR kit with Qiagen custon primers assays for mir-499 and mir-208. No-matrix PCR showed one-peak melting curve and the C(t) was 30, when in positive control C (t) was 34. I cheсked twice, so this is not because I changed the tubes. How it is possible?

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