Explore recombinant DNA technology through a comprehensive 12-hour video series covering essential topics such as DNA cloning, plasmid isolation, microarray techniques, and various blotting methods. Learn about PCR types, genomic library screening, and expression vectors. Dive into the construction and screening of genomic and cDNA libraries, including techniques like blue-white screening and colony hybridization. Understand the principles of real-time PCR, its various applications, and specialized PCR methods like nested, inverse, and hot start PCR. Examine different cloning vectors, including plasmids, cosmids, and artificial chromosomes. Gain insights into restriction enzymes, DNA methylation, and advanced cloning techniques like Topo cloning. Master the intricacies of molecular biology techniques essential for genetic engineering and biotechnology research.
Overview
Syllabus
DNA cloning.
Plasmid isolation protocol.
DNA microarray.
DNA ligase mechanism.
Role of alkalyne phosphaase in molecular cloning.
Role of linker and adaper DNA.
PCR types.
Genomic library screening.
PCR (polymerase chain reaction) in detail.
Southern blot protocol.
Western blot.
Northern blot.
Southern blot.
Expression vector.
Cloning vector.
Genomic library.
cDNA library.
PCR optimization | PCR technique.
Yeast artificial chromosome (YACs).
Plasmid as a cloning vector.
Cosmid vector.
Control of gene expression in pET vector.
Difference between genomic DNA library and cDNA library.
Construction of genomic library using yeast artificial chromosome.
Construction of genomic library using lamda phage.
Construction of genomic library using cosmids.
Blue white screening of DNA clones.
Using oligonucleotide probe to screen clones from genomic DNA library.
Replica plating.
Immunological screening (genomic library screening method).
Expression screening (genomic library screening).
Colony hybridization assay.
Synthesis of probes for Southern blotting.
Southern Blotting overview.
Real time PCR.
Real time PCR overview.
Real time PCR using SYBR green probe.
Real time PCR using scorpion primer.
Nested PCR.
Inverse PCR.
Hot start PCR.
Colony PCR.
Real time PCR using taqman dye.
Real time PCR using molecular beacon.
Quantitative real time PCR (qPCR).
Expression vector, pET vector.
Expression vector, pBAD vector.
Multiplex PCR.
M13 vector, a type of viral vector for cloning.
Cloning vector (lamda phage vector).
Genomic library screening: differential screening.
Bacterial artifical chromosome (BACs).
Southern blotting applications.
Properties of restriction enzymes.
DNA methylation.
Topo cloning.
Restriction enzyme digestion (Isochizomer, neoschizomer and isocaudamer).
Difference between linker and adaptor DNA.
Star activity.
Error prone PCR.
Degenerate PCR.
Taught by
Shomu"s Biology
