On Tue, Jan 10, 2017 at 8:00 AM, CodeWarrior <code.w4rri0r@gmail.com> wrote:
The TdT thus bound is still only non covalently bound to the DNA though. Even if an extreem PH solvent capable of removing it can't be found a sufficiently hot solvent will denature the protien alowing it to unbind but leaving the DNA intact.
Similar methods were described earlier in the thread (such as from Nathan):
https://groups.google.com/d/msg/diybio/FXaT-MjJQYI/tdHZwXqeCgAJ
https://groups.google.com/d/msg/diybio/FXaT-MjJQYI/tdHZwXqeCgAJ
DNA synthesis chemistry using TdT:
http://2014.igem.org/Team:Cooper_Union/TdT_project
http://2015.igem.org/Team:Cooper_Union/DeNovoSynthesis
http://2014.igem.org/Team:Cooper_Union/TdT_project
http://2015.igem.org/Team:Cooper_Union/DeNovoSynthesis
There was some discussion about that team's use of TdT over here:
https://groups.google.com/d/msg/enzymaticsynthesis/DApMjXx8gS4/CmKEK9-vJlwJ
https://groups.google.com/d/msg/enzymaticsynthesis/DApMjXx8gS4/CmKEK9-vJlwJ
even earlier mumblings about TdT from Nathan can be found here:
http://gnusha.org/logs/2013-07-13.log
http://gnusha.org/logs/2014-12-26.log
http://gnusha.org/logs/2013-07-13.log
http://gnusha.org/logs/2014-12-26.log
e.g. from a search he asked me to do a few years ago, http://diyhpl.us/~bryan/irc/nmz787-nucleotide.log.txt
TdT is certainly a useful critter to work with. A bunch of TdT chimeras and mutants would be really useful to investigate. By the way, I think that another area worth considering is DNA assembly; we can somewhat reliably get 100 bp oligos, and constructing longer stretches is a pain in the butt--- if we could somehow make that process simpler (such as using yeast homologous recombination) then we wouldn't need long DNA synthesis methods in the first place.
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