DNA Assembly

소개

Provides specialized procedural knowledge for synthetic biology tasks, focusing on high-fidelity DNA assembly methods like Golden Gate and Gibson assembly. The skill ensures accuracy in primer design by enforcing mandatory structures—such as clamp sequences and recognition sites—while providing systematic verification strategies to avoid common errors in overhang selection, reading frame preservation, and melting temperature calculation. It serves as a comprehensive guide for researchers creating fusion proteins or assembling multiple DNA fragments using specific enzyme variants like BsaI-HFv2.

주요 기능

• Strategic 4bp overhang selection and validation for multi-fragment assembly
• 16 GitHub stars
• Standardized Tm calculation and binding region optimization guidelines
• Verification checklists for reading frames, stop codons, and internal restriction sites
• Comprehensive primer structure design for Type IIS enzymes including mandatory clamps
• Reusable patterns for sequence parsing and programmatic design validation

사용 사례

• Designing primers for complex Golden Gate multi-fragment assemblies
• Developing fusion proteins while maintaining reading frames and removing internal stop codons
• Validating assembly designs against manufacturer-specific enzyme requirements like BsaI-HFv2