Fledgling grad student, Joe Peon, is doing PCR and goes to the freezer to get th

Question

Fledgling grad student, Joe Peon, is doing PCR and goes to the freezer to get the reagents needed to set up the reaction. This overworked graduate student grabs dNTPs, 2 forward primers, enzyme buffer, and restriction enzyme TaqI. out of storage and sets up his reaction tube. He sets up 2 tubes:

1) the positive control with target DNA added

2) the test sample + target DNA. He adds all the reagents listed, in the amounts given in the protocol and sets up the PCR cycler to cycle a heat phase for 30 seconds and extend for 1 minute. The next morning, he ran the samples out in an agarose gel.

a.What, if anything will Joe Peon see on the gel?

b What mistakes were m.ade (there are numerous errors – name 3 specific errors)?

Explanation / Answer

And a: no plication will be observed because polymerase enzyme is lacking. He can get many bands if the template is sufficient enough because the restriction enzyme will cleave the fragment atultiple restriction sites.

Ans b: the taq polymerase is not added. A restriction enzyme was added which shouldn't have been added. The magnesium chloride is missing.

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