flowchart TD subgraph Track1["Gene Cloning"] RE["Restriction Enzymes
Cut DNA at
specific sequences"]:::cloneNode GEL1["Gel Electrophoresis
Separate fragments
by size"]:::sharedNode CLONE["DNA Cloning
Insert into vector"]:::cloneNode RDNA["Recombinant DNA
Gene joined
to plasmid"]:::cloneNode TRANS["Transform Bacteria
Introduce plasmid
into host cells"]:::cloneNode SCREEN["Screen Colonies
Identify cells
with target gene"]:::cloneNode end subgraph Track2["Gene Editing"] PCR["PCR
Amplify target
DNA region"]:::sharedNode CRISPR["CRISPR-Cas9 Design
Create guide RNA
for target site"]:::editNode DELIVER["Guide RNA + Cas9
Deliver to cells"]:::editNode REPAIR["DNA Repair Pathway
NHEJ or HDR"]:::editNode SEQ["DNA Sequencing
Verify the edit"]:::sharedNode end RE --> GEL1 GEL1 --> CLONE CLONE --> RDNA RDNA --> TRANS TRANS --> SCREEN PCR --> CRISPR CRISPR --> DELIVER DELIVER --> REPAIR REPAIR --> SEQ PCR -.->|"Amplify insert"| CLONE GEL1 -.->|"Verify edit size"| SEQ SCREEN -.->|"Sequence to confirm"| SEQ classDef cloneNode fill:#3498DB,stroke:#2471A3,stroke-width:2px,color:#fff,font-size:16px classDef editNode fill:#27AE60,stroke:#1E8449,stroke-width:2px,color:#fff,font-size:16px classDef sharedNode fill:#8E44AD,stroke:#6C3483,stroke-width:2px,color:#fff,font-size:16px linkStyle default stroke:#999,stroke-width:2px,font-size:16px style Track1 fill:#EBF5FB,stroke:#3498DB,stroke-width:2px,color:#2C3E50,font-size:16px style Track2 fill:#EAFAF1,stroke:#27AE60,stroke-width:2px,color:#2C3E50,font-size:16px