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Biotech_Cloning Workflow

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Typical Biotech/Cloning Workflow Cloning describes the processes used to create an exact genetic replica of another cell, tissue, or organism. Researchers use a Biotech/cloning Workflow like this one that documents the sequence of activities in the cloning processes. DNA Extraction Isolates the DNA from the host culture to generate a purified DNA sample. Tissue Harvesting PCR (or qPCR) Polymerase Chain Reaction amplifies your gene of interest in your DNA sample. Restriction Digestion Restriction digestion cuts your amplified DNA's ends at points specific to certain enzymes. Preps for ligation into plasmid or further targets gene of interest. Ligation Ligation glues the newly chopped pieces of DNA with compatible ends into one plasmid. Gel Electrophoresis Runs your DNA fragments on a matrix that separates them based on size (the larger the fragment, the slower it moves). It can be used as a QC step or to visualize the result. Transformation Transformation incorporates the DNA into the host's genome, typically bacterial so that the host can make millions of copies as it grows. Cell Culture Sequencing (NGS, Sanger) Determining the order and frequency of the four nucleotide bases (adenine, guanine, cytosine, and thymine) in a sequence. ELISA "Enzyme-Linked-ImmunoSorbent-As- say; used to determine the quantity of a protein, antibody, antigen, or other material in a sample. Proteomics The large scale study of proteins and the genetics behind their function. Blotting The transfer and immobilization of proteins onto a surface. Commonly used for the detection and identifica- tion of various proteins, antigens, or other macromolecules.

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