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EK ICE 5
Divya_4978
#1 Posted : Tuesday, June 30, 2020 11:14:29 PM
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Hello,

I was wondering if you could explain question 72 of passage 1


Thank you in advance.
INSTR_Katerina_102
#2 Posted : Thursday, July 02, 2020 4:38:43 PM
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Hi,

Can you clarify, is this the question in EK ICE 4 regarding duplex PCR with the RNA from the TP53 gene?

Thanks!
Divya_4978
#3 Posted : Friday, July 03, 2020 12:46:50 AM
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Yes sorry, question 72, ICE 4 passage 1

Thank you
INSTR_Katerina_102
#4 Posted : Saturday, July 04, 2020 3:03:11 AM
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Hi,

This is a bit of a tough question.

You have been given only the RNA of the TP53 gene as stated in the question stem(here the implication is mRNA). This mRNA has been spliced at this point, removing introns and promotor sequences starting transciption.

To do this Duplex PCR sequencing technique , you need to have double stranded DNA, so you would have to get your DNA from the reverse transcription of the mRNA provided.

Because exons 3 and 5 are expressed in the mRNA, it is likely that mutations in both can be easily detected in Duplex PCR, eliminating B and C. Similarly, the DNA you get from reverse transcription also has an untranslated region, meaning D is unlikely as well.

However, the AT regions of DNA upstream of the promoter usually refer to a TATA box, which is part of the promoter region that is spliced out of mRNA - because our source of information is mRNA, we would not be able to see this region and therefore cannot comment on its variation, suggesting that A is the correct answer.

Please let me know if this is unclear, I hope this helps.
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