OligoSim

Currently I am testing whether or not I can directly use oligos to clone with in GoldenGate. Because of methods like PaperClip and BASIC, and also use of direct cloning to make sgRNAs, I assume this answer is yes. Planned experiments-

Best duplex buffer? IDT duplex or NEB RS buffers more efficient?

BsaI sites or raw overhangs? Is including BsaI sites at the end of oligos increase the efficiency because there is no requirement for T4 PNK, or less efficient, because it sequesters BsaI?

Complex parts? Can I anneal and clone terminators with high secondary structure?

Efficiency vs normal parts? (self explanatory)

OligoSim Map

Also shout out to protoza noah, your comments have been inspirational

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