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Real Time-PCR probe design

Real Time-PCR probe design - Real-Time PCR and Quantitative PCR Forum

Real Time-PCR probe design - Real-Time PCR and Quantitative PCR Forum


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  #1  
Old 08-07-2009, 01:06 PM
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Default Real Time-PCR probe design



Dear All,

I have to design a probe for qPCR laying between a pair of primers that I know is working quite good (with sybr green). I have a couple of questions about it:

1) Would it be ok if there is no free space between the 3-end of the probe and the 5-end of the reverse primer?

2) The Tm difference between the primers pair and the probe is about 6C (primers ~62C; probe ~68C). Is it enough?

All the other conditions for the probe that I am aware of are more or less fulfilled: no repetitions of more that 3 nucleotides; nor G nor C at the 5-end; length: 28 nucleotides; low self complementarity.

What do you think about it?

Thank you all
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Old 03-22-2010, 08:00 PM
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Default Re: Real Time-PCR probe design

have you tries your new primer and probe?, did it work?, if not, you may try anti-sense probe that will make 3' of probe far from reverse primer
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design , probe , qpcr , qpcr probe design , real , timepcr


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