1. 1 SURVEY OF BIOCHEMISTRY Nucleic Acids continued… Amino Acids

2. 2 PRS Question Name the following compound 1.ATP 2.UTP 3.GTP 4.CTP

3. 3 PRS Question Thymine has a methyl group at which position? 1.C1 2.C3 3.C5 4.C1’

4. 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. 5 Genomic Library Cut Genomic DNA with restriction enzymes Insert fragments into vectors Culture cells Reference: Wikipedia

6. 6 Making cDNA Image from Biochemistry by Stryer

7. 7 Making cDNA Clones Image from Biochemistry by Stryer

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

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