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SURVEY OF BIOCHEMISTRY Nucleic Acids continued… Amino Acids

SURVEY OF BIOCHEMISTRY Nucleic Acids continued… Amino Acids. PRS Question. Name the following compound ATP UTP GTP CTP. PRS Question. Thymine has a methyl group at which position?. C1 C3 C5 C1’. Last Week…. Recombinant DNA Technology Vectors, Cloning and Libraries

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SURVEY OF BIOCHEMISTRY Nucleic Acids continued… Amino Acids

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  1. SURVEY OF BIOCHEMISTRYNucleic Acids continued…Amino Acids

  2. PRS Question • Name the following compound • ATP • UTP • GTP • CTP

  3. PRS Question • Thymine has a methyl group at which position? • C1 • C3 • C5 • C1’

  4. Last Week… • Recombinant DNA Technology • Vectors, Cloning and Libraries • Genomic vs. cDNA Library • How are libraries made? • PCR (for making copies of DNA) • How does PCR work? • What is required to conduct PCR?

  5. Reference: Wikipedia Genomic Library • Cut Genomic DNA with restriction enzymes • Insert fragments into vectors • Culture cells

  6. Making cDNA Image from Biochemistry by Stryer

  7. Making cDNA Clones Image from Biochemistryby Stryer

  8. 3 steps for each PCR cycle • Each PCR cycle includes: • A denaturation step (92-96oC) separates the two DNA strands. • A primer annealing step (40-75oC) which is a few degrees below the Tm of the primers. • A primer extension step (72oC) which is the optimal temperature for Taq DNA polymerase activity.

  9. Components for PCR KNOW THIS!! • Heat-stable DNA polymerase Thermus aquaticus Taq polymerase • Primers (DNA oligonucleotides) • Deoxynucleotides –dATP, dTTP, dCTP, dGTP • DNA template • Mg++, buffer components, and water

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