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direct seq


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3 replies to this topic

#1 epigenetics

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Posted 22 April 2009 - 08:13 AM

is there anybody who is working on direct sequencing of bisulfite modified sample, using Tag added primers.

Please let me know how much success you are getting?
thanks,

#2 methylnick

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Posted 22 April 2009 - 01:39 PM

I have recently heard ABI have an optimised protocol and will be publishing it very soon. From a reliable source, ABI themselves!

This is with the SOLiD platform.

Nick

#3 Davo

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Posted 22 April 2009 - 05:05 PM

I've had very much mixed success with direct sequencing of bisulfite PCR products. I didn't use labelled or tagged primers however, I just used the primers from PCR. Sequencing of some amplicons worked brilliantly, some I couldn't get to work no matter what I tried.

The work by ABI Nick mentions will be an interesting read.

#4 epigenetics

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Posted 23 April 2009 - 06:03 AM

I was trying to do Direct sequencing since couple of months, Not working.
so Recently i tried tagged primer and waiting for result.
If ABI one works, that would be the best when you have lot of samples and number of genes to look, like us.

So eagerly waiting for that.

Thanks.




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