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why not working msp primers

MSP primer trouble

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4 replies to this topic

#1 zyk

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Posted 20 May 2015 - 12:25 AM

Hi

 

I have 3 genes to study MS PCR  but I tried 5 times msp primer desingn with meth primer programs didn’t get any results. Afterward I designing my own primers without any programs, I tried different temperature and time; again failure.   My times is running out and I dont know what can ı do. These gene promoters are difficult for primer designing. I attach my sequences and my own primers file. Could you please help

 Thank you

Attached Files



#2 methylnick

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Posted 21 May 2015 - 04:52 PM

Hi zyk, how are you bisulfite converting your DNA? are you running the controls that are usually included in the kit?

 

cheers

 

Nick


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#3 zyk

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Posted 21 May 2015 - 10:18 PM

Hi

 

I converting DNA with EZ methylation gold kit (ZYMO). The kit not involved controls . Therefore I controlled bisufite converted DNA with MSP primers that I used before and they worked.

 

Thank you for your reply



#4 methylnick

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Posted 22 May 2015 - 01:25 PM

I would suggest you revisit primer design and use methprimer express from Thermo Fisher.


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#5 zyk

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Posted 23 May 2015 - 11:54 AM

Thank you i will try




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