How to calc Tm for Gibson primers?

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Hello, I want to separate two purified proteins that have almost the same weight. One is 16.8 kDa, the other is approx 10kDa. I use Tris\glycine buffers. Which gel percentage and running time...

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In PRIMER, is it possible to downweight certain taxa in multivariate community datasets to reflect uncertainty around identification??

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PACYC PCR not working after several attempts. Would anyone have any suggestions?

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How to calculate the annealing temperature of a primer pair?

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Anybody with an idea on how do design specific primers for detecting tomato resistance genes tm-1, TM-2 and Tm2^2?

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What is the ideal efficiency for a qPCR?

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MiRNA qpcr problem?

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Calculating volume of DNA required from DNA concentration?

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Has anyone used the Illumina ITS1 primer sets?

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Genescan gives 3 peaks instead of one?

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QPCR reaction efficiency with odd results what does this indicate?

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Kelly Murray

The Tm should be calculated based on the annealing sequence only.

Mert Sudagidan

Dear Mi, the following links can help you.

http://www6.appliedbiosystems.com/support/techtools/calc/

https://eu.idtdna.com/calc/analyzer

Matthew de Cruz

I agree with Kelly, the TM should be calculated on the annealing sequences. The overhangs shouldn't be taken into consideration.

Mi La

Thank you all!