Lec8W11 - Finish plasmids Review ligation results Predict...

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Finish plasmids Review ligation results Predict transformation results Alkaline phosphatase Bioinformatics
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Ligation that worked well Ligation that did not work well
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Transformation results? Colonies – yes or no If yes, more blue or white? T1 through T4 (Ligation L1-L4) T5 – linear pGEM(L5) T6 – water T7 – uncut pGEM
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Alkaline phosphatase If a plasmid is treated with alkaline phosphatase after cutting, it cannot religate. Remember that ligase requires both a 5’phosphate and a 3’OH – alkaline phosphatase leaves 5’OH
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Ligase will work if insert still has a 5’phospahte – means of generating only circular plasmid vectors with inserts – linear plasmids won’t transform
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Bioinformatics is a term used to describe the databases and computational tools used in biology, with an emphasis on molecular biology. A biological database is a large, organized body of persistent data, usually associated with computerized software designed to update, query, and retrieve components of the data stored within the system. Bioinformatics
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Bioinformatics databases There are many different types of databases: DNA databases (nucleotide sequence) Protein databases (sequence, structure) Organism specific databases There are three main nucleotide databases that form the International Nucleotide Sequence Database Collaboration : Genbank, Embl, DDBJ These databases exchange information routinely. Protein databases include Swiss-Prot, PDB, and sequences that were created from translations of coding regions in DNA sequences stored in GenBank.
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Genbank GenBank serves as an public access repository for all
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Lec8W11 - Finish plasmids Review ligation results Predict...

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