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BIO311C_F07_DNA cloning_1

BIO311C_F07_DNA cloning_1 - Cloning Pages 384-391...

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Cloning Pages 384-391, 411-420 (relevant parts)
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We will cover Cloning Stem cells
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Cloning Process of creating an identical copy of the original Molecular cloning Reproductive cloning
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Molecular cloning Making multiple identical copies of a particular gene/s of interest
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Components of molecular cloning DNA to be cloned Cloning Vectors Carry DNA inside cells and replicate Plasmids Bacteriophages or viruses Artificial chromosomes Bacteria Enzymes
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Cloning vectors DNA spread from E.coli
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Cloning vectors Plasmids Origin: bacteria Circular DNA distinct from bacterial chromosome (extra chromosomal) Able to replicate Have origin of replication sequences
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Enzymes in molecular cloning Restriction endonuclease Bacterial enzymes Cut DNA molecules at a limited number of specific DNA sequences, called restriction sites Form restriction fragments DNA ligase seals the bonds between restriction fragments
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Restriction enzymes Recognize a specific sequence of bases in dsDNA and cuts within within that sequence Called the Recognition Site - can be 4-8 Recognition Site - can be 4-8 nucleotides nucleotides Cuts large fragments of DNA into smaller pieces - called a restriction digest or restriction digest or digestion digestion
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Restriction Enzyme Sites Have Point of Symmetry Restriction Enzyme Sites Have Point of Symmetry make staggered cuts because the sequences they recognize are usually symmetrical Called Palindromes Palindromes Palindrome : Base sequence read 5’ 3’ on one strand is same as base sequence read from 5’ 3’ on complementary strand.
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