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Old 05-05-2011, 12:36 AM   #2
tonybolger
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Location: berlin

Join Date: Feb 2010
Posts: 156
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Quote:
Originally Posted by ychang View Post
Hi, every, I want to prepare a library for mate pair end seq, the special aspect of the library is that I want the length of the fragment as long as possible, say, 10kb or even 20kb. Do you think the circularization efficiency will reduce with the increase of the length?

Thanks!
You can push the illumina protocol to at least 12Kb, using the 10Kb semi-official protocol but the quality heads downwards.

One alternative is to use the 454 linker approach, but ligate illumina adapters on the result - this should work up to 20Kb, and can possibly be pushed up to 65Kb with some tweaks, according to this

Another alternative is the fosmid approach, detailed here

I would be interested to know if any of these approaches work for you
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