I am new to qPCR technique, recently i started working on a project my standards are from 6 million to 60 copy number for two genes. When i perform qPCR the CT values differs by (1-4 ct). There are no pipetting errors or contamination. Mol weight of plasmid 1 : 1,474,284.6g/mole and plasmid 2: 1,465,624.1 g/mole. Can someone please explain me the reason behind such differences between standards with same copy number.

Thank you in advance.

J

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