Biotechnology: Principles and Processes Part-5
Polymerase Chain Reaction
graph TB
PCR(Polymerase Chain Reaction)
PCR --> DNA[DNA Denaturation]
PCR --> Primer[Primer Annealing]
PCR --> Extension[Extension/elongation]
DNA --> T1[Temperature: 94-98°C]
Primer --> T2[Temperature: 50-65°C]
Extension --> T3[Temperature: 72°C]
T1 --> T1a[Breaks hydrogen bonds]
T2 --> T2a[Primers bind to target DNA]
T3 --> T3a[DNA polymerase synthesizes new DNA]
DNA
graph TD;
DNA{{DNA}}
DNA --> Nucleotides{{Nucleotides}}
DNA --> DoubleHelix{{Double Helix}}
DNA --> Replication{{Replication}}
Nucleotides --> Adenine{{Adenine}}
Nucleotides --> Thymine{{Thymine}}
Nucleotides --> Cytosine{{Cytosine}}
Nucleotides --> Guanine{{Guanine}}
DoubleHelix --> BasePairing{{Base Pairing}}
Replication --> SemiConservative{{Semi Conservative}}
Replication --> Enzymes{{Enzymes involved in DNA replication}}
DNA Synthesis
graph TB
A[DNA Synthesis]
A --> B[DNA Replication]
B --> C[Helicase]
B --> D[DNA Polymerase]
B --> E[Primase]
A --> F[DNA Repair]
F --> G[DNA Ligase]
F --> H[Exonuclease]
F --> I[Endonuclease]
A --> J[DNA Recombination]
J --> K[Homologous Recombination]
J --> L[Site-specific Recombination]
J --> M[Transposition]
Recuirments for PCR
graph TD;
PCR[Requirements for PCR];
PCR --> Primer[Primers];
PCR --> dNTPs[dNTPs];
PCR --> Buffer[Buffer];
PCR --> Taq[Taq Polymerase];
PCR --> Template[Template DNA];
PCR --> Machine[PCR Machine];
Primer --> Forward[Forward Primer];
Primer --> Reverse[Reverse Primer];
dNTPs --> Adenine[A];
dNTPs --> Thymine[T];
dNTPs --> Cytosine[C];
dNTPs --> Guanine[G];
Buffer --> pH[pH];
Buffer --> Mg[Mg++];
Taq --> Heat[Heat Resistance];
Template --> Source[Source of DNA];
Machine --> Cycles[Cycling Conditions];
Polymerase Chain Reaction Stages
graph TD;
PCR["Polymerase Chain Reaction Stages"];
PCR --> Denaturation;
PCR --> Annealing;
PCR --> Extension;
Denaturation["1. Denaturation: DNA strands separate"];
Annealing["2. Annealing: Primers bind to the DNA"];
Extension["3. Extension: New DNA is made by the Taq polymerase"];
Instrumentation: Thermal cycler
graph TD;
A[Instrumentation: Thermal cycler];
B[Definition];
C[Uses];
D[Types];
E[Components];
F[Working Principle];
G[Applications in Biology];
A-->B;
A-->C;
A-->D;
A-->E;
A-->F;
A-->G;
Analysis of PCR Reaction
graph TD;
A[Analysis of PCR Reaction];
B[PCR Basics];
C[PCR Components];
D[PCR Steps];
E[PCR Variations];
F[PCR Applications];
G[PCR Limitations];
A-->B;
A-->C;
A-->D;
A-->E;
A-->F;
A-->G;
B-->H{DNA};
B-->I{Primers};
C-->J{DNA Polymerase};
C-->K{dNTPs};
C-->L{Buffer};
D-->M[Denaturation];
D-->N[Annealing];
D-->O[Extension];
E-->P[Real-time PCR];
E-->Q[Reverse transcription PCR];
E-->R[Digital PCR];
F-->S[Genetic Testing];
F-->T[Forensics];
F-->U[Research];
G-->V[Errors];
G-->W[Sensitivity];
G-->X[Specificity];
Restriction Digestion and Ligation
graph TB
A[Restriction Digestion and Ligation]
A --> B[Restriction Digestion]
B --> C[Definition]
B --> D[Process]
B --> E[Applications]
A --> F[Ligation]
F --> G[Definition]
F --> H[Process]
F --> I[Applications]
