Cloning Strategy Pros/Cons
There are various factors to consider when choosing between these two cloning strategies: number of steps to completion, ease of linker switchability, time for ordering parts, etc.
Typical strategy
Pros: Fewer steps, no pEXPR internal cut site problem, can easily change promoter
Cons: High number of total reactions (each linker construct has both Golden Gate and LR), requires prompt arrival of both primers and linkers, trying new linkers involves more steps
Alternative strategy
Pros: Low number of total reactions, includes screening mechanism for determining whether linker insertion occurred, allows extra time for linkers to arrive, easily allows for trying different linkers
Cons: More steps, requires prompt arrival of primers, pEXPR BsaI site, must use BP reaction in order to change promoter
Typical Cloning Strategy
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Alternative Cloning Strategy
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