dna-assembly
DNA Assembly
Overview
This skill provides procedural knowledge for DNA assembly tasks, particularly Golden Gate assembly using Type IIS restriction enzymes. It addresses common pitfalls in primer design and provides verification strategies to ensure correct assembly designs.
Critical Requirements for Type IIS Restriction Enzyme Primers
Complete Primer Structure
When designing primers for Type IIS enzymes (BsaI, BsmBI, etc.), the complete structure must include ALL components:
[clamp ≥1bp]-[recognition site]-[spacer N]-[4bp overhang]-[binding region 15-45bp]
For BsaI-HFv2 specifically:
[clamp ≥1bp]-GGTCTC-N-[4bp overhang]-[binding region]
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