GMO Investigator Lab: I am lost on this. We are using 2 different master mixes:
ID: 102523 • Letter: G
Question
GMO Investigator Lab: I am lost on this. We are using 2 different master mixes: 1) 20 ul Plant MM (green) 2) 20 ul GMO MM (red)
1. In this experiment, you will add a PCR “Master Mix” to each sample. What sorts of ingredients must be in this mix to allow the PCR reaction to work? For each ingredient you identify, be sure to specify its function in the reaction. (For help with PCR, see your text.)
2. For each sample you test in lab this week, you will set two PCR reactions. One of these reactions, labeled “Plant”, amplifies regions of DNA found in all plants. What is the purpose of this reaction? What does it tell you about your experiment?
please help.
3. You will also perform PCR on two samples known to contain GMO’s (tubes 5 and 6)! Why are these two samples included in our work? What can they tell us about the results of our experiment?
Explanation / Answer
1. To run the PCR reaction following ingredients just be added to the master mix.
A. DNA template - which should have the region which have to be amplified.
B. DNA polymerase enzyme - or generally Taq polymerase is used in here because of it's stable nature at high temperature. This enzyme is meant for formation of new DNA strands.
C. DNA primers - forward and backward primers to amplify the gene in a sequence specific manner.
D. dNTPs - used by DNA Polymerase enzyme to synthesize new strand of DNA.
E. Buffer solution- to give optimal chemical environment for DNA Polymerase to work.
F. Bivalent cations such as magnesium or manganese.
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