Koch Lab:Protocols/Fork unzipping constructs - Revision history

Steven J. Koch at 22:20, 14 June 2008

Steven J. Koch — Sat, 14 Jun 2008 22:20:43 GMT

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Steven J. Koch at 22:08, 14 June 2008

Steven J. Koch — Sat, 14 Jun 2008 22:08:11 GMT

←Older revision		Revision as of 22:08, 14 June 2008
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	==Principle==		==Principle==
	'''More details to come''' (Please send a note if you'd like us to fill in details sooner.)		'''More details to come''' (Please send a note if you'd like us to fill in details sooner.)
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	Unzipping forks can incorporate protein binding sites directly, or a cohesive end for ligation of long DNA unzipping fragments.		Unzipping forks can incorporate protein binding sites directly, or a cohesive end for ligation of long DNA unzipping fragments.
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Steven J. Koch: New page: ==Principle== '''More details to come''' (Please send a note if you'd like us to fill in details sooner.) DNA tethers suitable for short unzipping can easily be formed by hybridizing two...

Steven J. Koch — Sat, 14 Jun 2008 22:07:21 GMT

New page: ==Principle== '''More details to come''' (Please send a note if you'd like us to fill in details sooner.) DNA tethers suitable for short unzipping can easily be formed by hybridizing two...

New page

==Principle==
'''More details to come''' (Please send a note if you'd like us to fill in details sooner.)

DNA tethers suitable for short unzipping can easily be formed by hybridizing two DNA oligonucleotides with partial complementarity, and partial non-complementarity -- forming a fork construct. The non-complementary parts are best formed from poly(dT) stretches. The initial length of the tether will be the length of the two poly(dT) stretches, and due to limitations of synthesis length, will be shorter than in the [[Koch Lab:Protocols/Unzipping constructs|anchoring segment method]]. Among many challenges, one you need to look out for is ionic repulsion between microspheres and surface, which can inhibit tether formation.

Unzipping forks can incorporate protein binding sites directly, or a cohesive end for ligation of long DNA unzipping fragments.