1. You added a PCR “Master Mix” to each sample. List all of the ingredients that must be in this...

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1. You added a PCR “Master Mix” to each sample. List all of the ingredients that must be in this Master Mix to allow the PCR reaction to work. (Hint- Think, does Master Mix contain DNA?) For each ingredient you identify, be sure to specify its specific function in the reaction.2.Our PCR reaction will run for 40 cycles. In each cycle of PCR, there are three steps: Denature, Anneal and Extension. A) For each step, state the specific, required temperature and ALSO, explain B) what that specific temperature achieves.3.If human DNA polymerase were used in this PCR reaction, instead of Taq Polymerase, what temperature do you predict would be used for the Extension phase of the reaction? Why?4.Consider two parents. One parent has 5 repeats at the TH01 locus on both of her homologous chromosomes. The other parent has 10 repeats on both of his.

If you were provided with DNA from each of these individuals, and performed a PCR with primers specific to each end of the TH01 region, how long would the fragment you generated be for each of the parents? (Ignore the length of the primer in your calculations!
If you were provided with DNA from a child of theirs, what would you expect the results of your PCR analysis to look like? (That is, what size fragments would you generate?) Explain how you are able to make this prediction!
Now consider a crime scene in which two offspring from these same parents (siblings!) are both suspects. Could a DNA profile based on just this TH01 region determine which sibling had committed the crime? Why or why not? If not, how might you improve your profiling to be able to distinguish these two individuals?

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Bhawna · Accepted Answer

1. PCR master mix is prepared to perform the PCR by adding the template in them. The master mix have all the reagents required to perform the Polymerase Chain Reaction and these should be present in optimal conditions. The reagents used for PCR are:
dNTPs: These are the deoxynucleoside phosphates, they are required for DNA to extend. It consist of all the nucleosides such as adenine, thymine, cytosine and guanine. 
Buffer and cofactors: They are required in the mixture to provide optimum conditions for the process to take place. The buffer provide the optimum condition for the polymerase reaction to perform. The main co factor used is magnesium as it help in formation of phosphodiester bond. 
Primers: They are short oligonucleotides which recognize and then bind with the template DNA. These are the sequence which will be amplified in the whole process. Two type of primers are used,

1. You added a PCR “Master Mix” to each sample. List all of the ingredients that must be in this Master Mix to allow the PCR reaction to work. (Hint- Think, does Master Mix contain DNA?) For each...

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