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Multiplex PCR with Syber Green I

Multiplex PCR with Syber Green I - Real-Time PCR and Quantitative PCR Forum

Multiplex PCR with Syber Green I - Real-Time PCR and Quantitative PCR Forum


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  #11  
Old 11-19-2008, 07:12 PM
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Default Re: Multiplex PCR with Syber Green I

I agree, HRM could be the right thing there.
I don't know yet to what extent I will have better resolution comparing to SYBR Green I, but I'm going to have a try soon.
Does the resolution obtained using HRM also depend on the software offered by the machine producer?
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Old 06-15-2009, 02:59 PM
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Default Re: Multiplex PCR with Syber Green I

I would agree with Chovek as well. I my self is doing a multiplex PCR using SYBR and was able to optimize the method. However, working with close range Tms multiplexing becomes difficult. Chovek could you please provide me that study using the high-resolution melting curve?
thanks
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Old 06-16-2009, 03:06 PM
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Default Re: Multiplex PCR with Syber Green I

hi all,
may be could be useful try with saturating dyes, like EVA green and LCGreen in order to better define the Tm and shape of curves. Personally I'm tring to differentiate between three amplicons with a gap of 1c without a dedicated HMR Real Time system and it seems to work.
Does anibody use HR-1 from Idaho Technology?
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