Biotechnology Notes

Biotechnology overview

DNA modifying Enzymes Cutting and Joining DNA molecules, Genetic Engineering

       Restriction endonuclease - RE 1, RE 2, Factors that Influence Restriction Enzyme Activity, Enzymes used in R-DNA technology, DNA Methyltransferases

        Biology and Activity of Restriction Endonucleases, Cutting DNA with Restriction Endonucleases,

        Restriction Mapping

        Joining DNA molecules - DNA Ligation

        Nucleases: DNase and RNase, DNA Ligase, Alkaline Phosphatase, Polynucleotide Kinase, Alkaline Phosphatase

        DNA Polymerases

        E. coli DNA Polymerase I , Klenow Fragment of E. coli DNA Polymerase I

        T4 DNA Polymerase , T7 DNA Polymerase

        Terminal Transferase

        Thermostable DNA Polymerases (Taq, Pfu, Vent, etc.)

        Reverse Transcriptases

        Bacteriophage RNA Polymerases

        Adaptors, Linker, Homopolymer Tailing, Engineering DNA ends,

Plasmids and Vectors -

        Plasmids, Plasmids, Plasmid copy number, Extrachromosomal Elements, Bacterial plasmids and DNA transformation,

        Vectors, Vector Construction, Cloning vector and Expression vector

        Prokaryotic Expression Vectors 1, PE vectors 2, High-capacity vectors

        pBR322, pUC, Bacteriophage Lambda Vectors,

        Bacteriophage M13, M13mp Vectors, λ phage, Cosmids, λ cosmids

        P1 phage, Cosmids, BACs, PACs, Yeast Artificial Chromosomes (YACs)

        Virulent bacteriophage methods

Gene Cloning

        Basic cloning technique, Additional cloning methods, Cloning PCR Products, Cloning Genes,

        Humans Cloning Ethical issues

Cloning Strategies

        Libraries, M13 Phage Display Libraries

        Cloning Genomic DNA - Genomic DNA Libraries, Gene Libraries

        cDNA Cloning - cDNA, cDNA Libraries

        Screening -        

Sequencing

        DNA sequencing, DNA Sequence Analysis, Sequencing Assembly, Genome Annotation

Mutagenesis

        Site Directed Mutagenesis, 1

Gene Transfer

        Transgenic plants, Somatic cell genetics and transgenics,

Genetic Engineering

        Application, Combinatorial genetic engineering, Role of Microorganisms in Genetic Engineering

Recombinant DNA - re combining of two or more DNA molecules

        rDNA, Selectable Markers & Markers for Screening

        Gene function transmission (conjugation transformation transduction)

            1) isolation identification of desired gene

            2) insertion of desired gene in suitable Vector

            3) insertion of this vector to suitable host/organism

            4) selection of the transformed host cell

            5) multiplication/expression of the desired gene

Molecular Markers

        Markers, Marker Technology

        RAPD, RAPD's, Arbitrary-Primed PCR (Randomly Amplified Polymorphic DNA)

        VNTR's, AFLP's, RFLP's, SSCP's, RAPD

Basic Techniques of Biotechnology

        DNA Techniques -DNA Detection and Hybridization, DNA diagnostics, Primers

        Nucleic acid blotting - Southerns, Northerns, Westerns, & Cloning Molecular Searching Techniques

        Southern blotting, Northern blotting, Western blotting, Eastern blotting

        Polymerase chain reaction - PCR and Thermalcycling,  thermalcycler files

        RT-PCR, Long accurate PCR (LA-PCR), Real-time quantitive PCR

        Monoclonal Antibody Technology, Antibody methods

        SELEX (Selective Evolution of Ligands by Exponential Enrichment)

        Molecular Imprinting,

Gene Therapy

        Gene therapy

Nanotechnology and Microarray

        Chips and microarrays, Microarray, Microarray II

 

Nucleic acids and polymerization

Finding the gene that encodes a purified protein, Temperate bacteriophage methods,

Potential Term Paper Topics for Biotech