[DIYbio] A hypothetical protocol for DIYBIO DNA synthesis

My apologies for repeatedly flogging a dead horse of a thread but a new idea occurred to me this morning. If it could be made to work it would eliminate the need for special blocked dNTPs. Supose TdT could be modified to cause it to bind very tightly to DNA ends. So tightly that after nucliotide addition instead of advancing along the DNA strand it stays stubbornly in place blocking further addition. In the case the the dNTP no longer needs to be reversibly blocked as the TdT that has added a nucliotide now remains bound blocking further additions.

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.

One could invision a sequence in which one fist adds a modified TdT with a dNTP mix causing the majority of DNA strands to elongate. The solid suport would then be washed removing free TdT and dNTP. Then exposed to a solvent to denature / unbind TdT blocking strand elangation in preperation for further additions.

Does that sound viable? The question in my mind is how dificult might it be to protien engenear TdT incapable of progressing along or unbinding from a DNA strand end? I've no facilities to produce or test TdT mutants in a lab at the moment but given time I may be able to investigate the matter in silico.

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