Name: MACDONALD CHIEMEZIE MBAGWU
Date: 15/04/2024
Course: MOLECULAR MEDICICINE AND MEDICAL GENETICS
ASSIGNMENT.
School: VICTORIA UNIVERSITY OF BARBADOS
A. POLYMERASE CHAIN REACTION
Overview:
● Goal: Amplify small quantities of DNA
● Clinical relevance in modern medicine and clinical genetics: Diagnostic tool (ex. HIV, herpes
encephalitis).
Process:
● Denaturation: Heat DNA to 95°C to separate strands
● Annealing: Cool sample to 55°C
- Add DNA primers
- Add heat-stable DNA polymerase
- Add deoxynucleotide triphosphates (dNTP)
- Taq polymerase: Heat-stable DNA polymerase
- DNA primers anneal to desired portion of DNA strand
- Every PCR uses 2 primers to flank the desired region of transcription
● Elongation: Increase temperature to 72°C
- DNA polymerase adds dNTPs to DNA primer
●This process is repeated many times → Amplify DNA
● PCR results can be analyzed via gel electrophoresis
Reverse Transcriptase PCR:
●mRNA → cDNA → PCR
● Reverse transcriptase enzyme added prior to PCR
A1. CRISPR/Cas9
Overview:
● Goal: Gene editing (deletions, mutations, or insertions)
● Use: Genetic manipulation, cancer research, viral research, genome study
Process:
● Cas9: Bacterial endonuclease used to locate and cleave host DNA
