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Laboratory : Unit 4: purify amplicon  ligate to plasmid  transform; agarose gel (pages 84-86) Lecture : topoisomerase-mediated ligation; lacZ & alpha complementation In-Class Writing : edit sequences; Unit 3 (56-61) Read : Day, Chapter 27; manual 71-100, 213

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  1. Laboratory: Unit 4: purify amplicon  ligate to plasmid  transform; agarose gel (pages 84-86) Lecture: topoisomerase-mediated ligation; lacZ & alpha complementation In-ClassWriting: edit sequences; Unit 3 (56-61) Read: Day, Chapter 27; manual 71-100, 213 HandIn: editorial (pages xlii, section E & 96) DueNext Class: draft of lab report 3 (pp. 109-115)

  2. untemplated A

  3. Alpha complementation: Used in many vectors Distinguish E. coli with: empty vector from vector + foreign DNA

  4. Plasmid has: lac promoter/operator restriction sites for cloning 5' end of lacZ gene LacZ α-peptide

  5. Plasmid (NO insert)  LacZ α E. coli LacZ ω Lac+(blue colonies) Plasmid + foreign DNA NO LacZ α E. coli LacZ ω  Lac- (white colonies)

  6. X-Gal

  7. indigo

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