Biotechnology Masterclass: DNA, PCR, Gene Therapy & AI

Isolate and purify the target DNA, digest with restriction enzymes to create cohesive ends, ligate into a plasmid, transform bacteria, form a DNA library, and identify the desired clone.

Explore recombinant DNA technology and cloning to generate quantities of DNA by isolation, restriction enzyme cutting, ligation into cloning vectors, and host transfer for genetic mapping, sequencing, and PCR.

Restriction endonucleases cleave DNA at specific 4–6 nucleotide recognition sequences, creating sticky ends. These fragments with complementary sticky ends on antiparallel strands can be joined by DNA ligase.

Biotechnology today and yesterday covers how restriction enzymes recognize palindromic sites, cleave DNA to create blunt or sticky ends, and enable recombinant DNA cloning; bacteria defend against viruses using methylation.

Explore gene vectors and cloning vectors such as plasmids, phage lambda, cosmids, shuttle vectors, and yeast or bacterial artificial chromosomes, with origins, restriction sites, and selectable markers.

Discover cloning vectors, including lambda phage, cosmids, BAC and YAC, their replication cycles, packaging, and use in transferring large DNA fragments across hosts.

Develop genomic libraries by digesting genomes at restriction sites, cloning overlapping fragments into plasmid vectors in E. coli, followed by screening to identify positive clones.

Analyze restriction enzyme sites and map their positions using agarose gel electrophoresis with ethidium bromide, highlighting restriction fragment length polymorphism applications in forensics and gene analysis.

Use PCR to amplify DNA segments from a small sample, employing Taq polymerase, primers, and deoxyribonucleotides through cycles in a thermal cycler, identifying microorganisms and linking crime DNA.

Explore the polymerase chain reaction by starting with a template and primer, adding nucleotides, denaturing the double strand at 96°C, annealing at 56°C, and extending to repeat the cycle.

Microarrays use oligonucleotide probes on a solid surface to identify organisms and genes by hybridization, with laser-scanned fluorescence indicating probe hybridization.

Analyze genomes with dna microarrays and microchip arrays to measure expression from dna to rna to protein, explore bioinformatics predictions, and study regulatory gene networks across model organisms.

Explore genechip technology as a core topic in biotechnology today and yesterday, highlighting its role in the field.

Explore microarray technology to study gene interactions and regulatory networks, using fluorescently labeled cDNA on glass slides, hybridization, scanning, and clustering to reveal gene expression patterns.

Explore microarray analysis for human melanoma and breast carcinoma, isolating single cells, converting to labeled material, applying to the chip, and scanning oligo-hybridized DNA to detect activation states.

Explore DNA sequencing methods, including chemical Maxam-Gilbert and enzymatic Sanger sequencing, and learn how gel electrophoresis, labeling, and automation reveal nucleotide order.

Explore dna sequencing using a primer and dna polymerase with nucleotides, where termination creates fragments detected by polyacrylamide gel electrophoresis and laser-detected colored bands.

Explore Fred Sanger's dna sequencing method using a primer and template to synthesize a strand, with dideoxynucleotide terminators and gel electrophoresis to read the sequence from bottom to top.

Explore gel electrophoresis to separate DNA fragments by size using an agarose gel and electric current, visualized with ethidium bromide staining and a size marker.

Review the development of DNA sequencing methods from first to third generation, highlighting second generation platforms like 454 and Illumina, and Oxford Nanopore Technologies with bioinformatics to reduce errors.

Explore DNA sequencing workflows, from denatured double and single strands with primer annealing, through PCR amplification, polyacrylamide gel analysis, and software-based interpretation of sequencing data.

Use modified viruses as vectors to insert healthy genes into host cells, enabling gene therapy. Deliver the gene via an adenovirus vector to correct mutations and restore protein function.

Gene therapy treats inherited and acquired diseases such as cancer, AIDS, diabetes, Parkinson's disease, and cystic fibrosis by replacing defective genes using viral vectors, liposomes, or electroporation.

Compare viral vectors—retroviruses, adenoviruses, AAV, and HSV—with non-viral liposomes and direct injection methods for receptor-mediated endocytosis and internalization into target cells.

Learn how monoclonal antibodies target specific disease antigens, including cancers and infectious or autoimmune diseases, by hybridoma technology that fuses B lymphocytes with myeloma cells to create stable antibody clones.

Explore transgenesis through genetic modification and model organisms, examining gene knockin/knockout, embryo manipulation, nuclear transfer, and production of therapeutic proteins.