Hi all.
I am new to real time PCR.
Recently, I am doing real tine PCR on a plasmid containing a mutated gene. Then I use 2ng/ul plasmid to do a real time pcr using SYBgreen. However, I get a Ct value of 6 . I just wonder whether Ct is too small. And is it bad or does it have any implication if the Ct value is too small?
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Trof
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Posted 24 August 2011 - 11:17 PM
Yes, 6 is really too small. You should dilute your plasmid at least 1000x to get values between 15 and 30, that are suitable for quantification.
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