Description
DNA extraction: lyse cells → add phenol-chloroform (denatures proteins, partitions into organic phase) → centrifuge → aqueous phase contains DNA → ethanol precipitate → wash → resuspend. Yields pure DNA for downstream use.
Protocol Flowchart
graph TD
A[Cell pellet or tissue] --> Lyse[Lyse in buffer + protease]
B[Phenol-chloroform] --> AND1{Lysate AND
Phenol-chloroform?} Lyse --> AND1 AND1 --> PC[Add phenol-chloroform 1:1] PC --> Mix[Vortex and mix] Mix --> Spin[Centrifuge to separate phases] Spin --> Aq[Collect aqueous upper phase] Aq --> EtOH[Add 2× vol ethanol] EtOH --> Precip[Precipitate DNA] Precip --> Spin2[Centrifuge to pellet] Spin2 --> Wash[Wash pellet with 70% ethanol] Wash --> Dry[Air dry pellet] Dry --> Resus[Resuspend in TE or water] Resus --> QC[Quantify by spectrophotometry] style A fill:#ff6b6b,stroke:#c92a2a,color:#fff style B fill:#ff6b6b,stroke:#c92a2a,color:#fff style Lyse fill:#51cf66,stroke:#2f9e44,color:#fff style PC fill:#51cf66,stroke:#2f9e44,color:#fff style Mix fill:#51cf66,stroke:#2f9e44,color:#fff style Spin fill:#51cf66,stroke:#2f9e44,color:#fff style Aq fill:#74c0fc,stroke:#1971c2,color:#fff style EtOH fill:#51cf66,stroke:#2f9e44,color:#fff style Precip fill:#74c0fc,stroke:#1971c2,color:#fff style Spin2 fill:#51cf66,stroke:#2f9e44,color:#fff style Wash fill:#51cf66,stroke:#2f9e44,color:#fff style Dry fill:#74c0fc,stroke:#1971c2,color:#fff style Resus fill:#51cf66,stroke:#2f9e44,color:#fff style QC fill:#b197fc,stroke:#7950f2,color:#fff style AND1 fill:#b4b4dc,stroke:#8b8bc4,color:#000
Phenol-chloroform?} Lyse --> AND1 AND1 --> PC[Add phenol-chloroform 1:1] PC --> Mix[Vortex and mix] Mix --> Spin[Centrifuge to separate phases] Spin --> Aq[Collect aqueous upper phase] Aq --> EtOH[Add 2× vol ethanol] EtOH --> Precip[Precipitate DNA] Precip --> Spin2[Centrifuge to pellet] Spin2 --> Wash[Wash pellet with 70% ethanol] Wash --> Dry[Air dry pellet] Dry --> Resus[Resuspend in TE or water] Resus --> QC[Quantify by spectrophotometry] style A fill:#ff6b6b,stroke:#c92a2a,color:#fff style B fill:#ff6b6b,stroke:#c92a2a,color:#fff style Lyse fill:#51cf66,stroke:#2f9e44,color:#fff style PC fill:#51cf66,stroke:#2f9e44,color:#fff style Mix fill:#51cf66,stroke:#2f9e44,color:#fff style Spin fill:#51cf66,stroke:#2f9e44,color:#fff style Aq fill:#74c0fc,stroke:#1971c2,color:#fff style EtOH fill:#51cf66,stroke:#2f9e44,color:#fff style Precip fill:#74c0fc,stroke:#1971c2,color:#fff style Spin2 fill:#51cf66,stroke:#2f9e44,color:#fff style Wash fill:#51cf66,stroke:#2f9e44,color:#fff style Dry fill:#74c0fc,stroke:#1971c2,color:#fff style Resus fill:#51cf66,stroke:#2f9e44,color:#fff style QC fill:#b197fc,stroke:#7950f2,color:#fff style AND1 fill:#b4b4dc,stroke:#8b8bc4,color:#000
Color Scheme (6-Color System)
Red
Triggers & Inputs
Triggers & Inputs
Yellow
Structures & Objects
Structures & Objects
Green
Processing & Operations
Processing & Operations
Blue
Intermediates & States
Intermediates & States
Violet
Products & Outputs
Products & Outputs
Lavender
Decision Diamonds (OR/AND)
Decision Diamonds (OR/AND)
▼📊 Scientific Accuracy
This flowchart is based on verified molecular biology protocols from peer-reviewed literature. Phenol-chloroform extraction and precipitation steps have been validated against primary sources.
▼📚 Sources & Citations
Sambrook J, Russell DW. Molecular Cloning. 3rd ed. Cold Spring Harbor. 2001.Standard reference for DNA extraction.
Green MR, Sambrook J. Molecular Cloning. 4th ed. Cold Spring Harbor. 2012.
▼📋 Metadata
| Process ID | assay_protocols-dna-extraction |
| Created | 2026-03-15 |
| Verified | ✅ Yes |
| Last Updated | 2026-03-15 |
▼Process Statistics
Nodes: 15 | Edges: 15 | OR Gates: 0 | AND Gates: 1