PCR basics

What is the purpose of PCR?

• To produce millions of copies of DNA.
• To produce millions of copies of a specific region of DNA
• To add nucleotides to a DNA sequence
• To watch polymerase work.

What is the function of gel electrophoresis?

• To separate DNA fragments based on size.

• To amplify a specific region of DNA.

• To add nucleotides to a DNA sequence.

• To observe DNA replication.

What is the role of DNA polymerase in PCR?

• To separate DNA fragments based on size.

• To amplify a specific region of DNA.

• To cut DNA at specific recognition sites.

• To add nucleotides to a DNA sequence.

What is the role of DNA polymerase in PCR?

• To separate DNA fragments based on size.

• To amplify a specific region of DNA.

• To cut DNA at specific recognition sites.

• To add nucleotides to a DNA sequence.

Why do we use PCR for SSR-based genotyping in both animals and plants?

• To select among specific sites in the nuclear genome

• To selectively amplify organelles' DNA

• To test the quality of DNA prior to genotyping

• To size the alleles corresponding to each marker

Which one of the following is NOT a PCR mix?

• Buffer 5X, dNTPs, Hind III, F-Primer+R-Primer, MgCl2, Taq-DNA POL,

  template DNA.

• Buffer 5X, dNTPs, F-Primer+R-Primer, MgCl2, Taq-DNA POL,

  template DNA

• Buffer 5X, dNTPs, F-Primer+R-Primer, MgCl2, RNA ase,

  template DNA

• Buffer TAE 1X, dNTPs, ,F-Primer+R-Primer, MgCl2, Taq-DNA POL,

  template DNA

Establish a PCR thermal cycling scheme to amplify a 1000 bp target in a commercial plasmid vector used for cloning.

Switch to an amplification scheme to amplify an SSR marker mapping in a nuclear genome of Triticum aestivum (bread wheat)

What about SSR amplification of a nuclear pig genome? Would you rather use schem 1 or 2? Explain your ideas.

Agarose gel electrophoresis is NOT suitable for:

• sequencing

• checking DNA quantity

• Checking for RNA traces in a total DNA extraction

• sizing the alleles used to reconstruct plant and animal identity