This course on Recombinant DNA technology aims to teach students the skills needed for DNA cloning, plasmid isolation, DNA microarray, PCR techniques, genomic and cDNA library construction, Southern blotting, Western blotting, Northern blotting, and gene expression control using various vectors. The teaching method includes detailed explanations, protocols, and hands-on techniques. The course is intended for individuals interested in molecular biology, genetic engineering, biotechnology, or related fields.
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
