survey of biochemistry nucleic acids continued… amino acids

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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 - PowerPoint PPT Presentation

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Page 1: SURVEY OF BIOCHEMISTRY Nucleic Acids continued… Amino Acids

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SURVEY OF BIOCHEMISTRY

Nucleic Acids continued…Amino Acids

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PRS Question

• Name the following compound

1. ATP2. UTP3. GTP 4. CTP

Page 3: SURVEY OF BIOCHEMISTRY Nucleic Acids continued… Amino Acids

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PRS Question

• Thymine has a methyl group at which position?

1. C12. C33. C54. C1’

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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?

Page 5: SURVEY OF BIOCHEMISTRY Nucleic Acids continued… Amino Acids

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Genomic Library

• Cut Genomic DNA with restriction enzymes

• Insert fragments into vectors• Culture cells

Reference: Wikipedia

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Making cDNA

Image from Biochemistry by Stryer

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Making cDNA Clones

Image from Biochemistry

by Stryer

Page 8: SURVEY OF BIOCHEMISTRY Nucleic Acids continued… Amino Acids

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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.

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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

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