Determine which allele was mutated second.

You are a molecular pathologist resident who has just been given a large retinoblastoma collection and have been asked to categorize the collection into familial and non-familial cases. You have also been ‘asked’ to comment on the mechanism that took place in mutating the second allele. There is good news and bad news.
The bad news: someone deleted the files linking the family history to each sample.
The good news: for each retinoblastoma case, you have a single tumour sample and its matched normal tissue from each patient, meaning all the samples are paired.
A) Describe what evidence you would seek to determine whether or not you are looking at a familial vs sporadic case of RB.
B) Is there a situation where you could definitively determine which allele was mutated second? If so, how? If not, why not?
C) For tumors in the non-familial group what are the potential mechanisms for loss of the second allele.
D) Would familial RB be a candidate for gene therapy? Please give a short explanation why or why not.

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