75. Methods of DNA analysis. Recombinant DNA in Medicine. Single Nucleotide Polymorphism. Polymerase Chain Reaction (PCR) as a Diagnostic Tool.
Covers the main laboratory methods for analysing DNA, from restriction fragment analysis and blotting techniques to sequencing, FISH, microarrays and CRISPR-Cas9. It then outlines recombinant DNA uses in medicine, single nucleotide polymorphisms with coagulation examples, and PCR's steps, reagents and diagnostic applications, including real-time PCR and RT-PCR.
Topic 75 of 76 in the MU Pleven syllabus for Biochemistry (Medicine, Year 2). The free AceMed note covers it in 5 sections, about 357 words (3 min of reading), with 20 flashcards.
In this note
Methods of DNA analysis. Recombinant DNA in Medicine. Single Nucleotide Polymorphism.
Methods of DNA Analysis
Recombinant DNA in Medicine
Single Nucleotide Polymorphism (SNP)
Polymerase Chain Reaction (PCR)
Methods of DNA Analysis
Restriction Fragment Analysis: DNA is cut by restriction endonucleases at specific sequences; resulting fragments are separated by gel electrophoresis to compare patterns.
Southern Blotting: Detects specific DNA sequences. DNA fragments are separated, transferred to a membrane, and probed with labeled complementary DNA.
Northern Blotting: Same concept but detects RNA to evaluate gene expression.
Western Blotting: Detects proteins using antibodies.
DNA Sequencing: Determines the precise nucleotide order (Sanger sequencing or next-generation sequencing methods).
Fluorescence In Situ Hybridization (FISH): Uses fluorescent probes to detect gene location or chromosomal abnormalities directly on cells.
Microarray Analysis: Measures expression of thousands of genes simultaneously using complementary probes on chips.
CRISPR-Cas9 Based Analysis: Can identify and modify specific DNA sequences with high accuracy.
Recombinant DNA in Medicine
Recombinant DNA involves joining DNA fragments from different sources to create new genetic combinations.
Used to produce therapeutic proteins such as insulin, growth hormone, clotting factor VIII, erythropoietin.
Gene therapy uses viral or non-viral vectors to deliver normal genes into cells with defective ones.
