Hey,
I`m trying to design a RT-PCR from scratch. I`ve designed my primers and
have their Tm. But if I`m going to test them in RT-PCR how do I know which program I have to use?
I`m talking about annealling temperature, number of cycli...??
Are there just standard parameters you use and you just fill in your Tm for your primers?
Thanks
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1 reply to this topic
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Curtis
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Posted 07 January 2010 - 01:42 AM
you need to run gradient to determine the exact annealing temp.
you can also buy kits for RT-PCR from various companies. the protocols are in the kits or you can even find them online. there are two types of RT-PCR; one-step and two-step.
it would be better if you could have a look at protocol-online's main page for more info.
you can also buy kits for RT-PCR from various companies. the protocols are in the kits or you can even find them online. there are two types of RT-PCR; one-step and two-step.
it would be better if you could have a look at protocol-online's main page for more info.
