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Trouble with primers of BSP - (Feb/20/2006 )

Hello, everybody! I'm new about methylation study and I am trying to do bisulfite sequencing. I try more time to amplify the sequence, but nothing. My primers are designed by MethPrimer. I guess maybe my primers are poor. People suggest some software online, I want to know which one is good. And if I should design by myself? Another question is that the primer should be synthesised in special company?

I look forward to your help.



I find it easier to design primer's myself.

I have had mixed results using methprimer I have to say. I think I have posted notes on primer design previously.

There is no need for special sythesis of primers, I usually have mine desalted (the default purification) and they work fine.

If you would like some notes on primer design I will post them shortly.



Your are friendly to me.
Yes I would like some notes on primer design from you!
Today I use methprimer to design a pair of BSP primers, but there is non-specific strip and smear, I don't know how to do. Can you help me? Thank you!



Another question, how do you design MSP primers by yourself?

I want to find if methylation degree are different in pig tissues. Frist I want to use MSP primers to find if there are differences. And if there are, I will use BSP primers to sequence or use DHPLC to detect the methylation degree. Do you think this is good?

Thank you!