6) To set up the PCR reaction, we added 20 uL of extracted DNA to 20 ul of There

Question

6) To set up the PCR reaction, we added 20 uL of extracted DNA to 20 ul of Therefore, we made a 1: 2 dilution of the master mix. 7) Jack is looking for GMO food, and sets up the following PCR reactions (as we di he gets a strange result (in the table below). -Indicate which results gave an expected vs an unexpected result in the last row of the cha s we did in class). Howe rt GMO positive GMO positive Template DNA GMO Free GMO Free Test Food Test Food Primers Plant GMO sequence GMO sequence Plant GMO Plant sequence sequence sequence sequence DNAeled DNAdRNA.oeled negative DNAcid Expected orunype ic te 6) Give a possible explanation for why his control reactions came out incorrectly (many answers are ? ossible, you may be creative). H PCR rcaction ncorrect

Explanation / Answer

6) Here we have added 20uL of DNA to 20uL of master mix that means the resulting solution is 2X diluted. In terms of ratio it's 1:1 dilution of the master mix

7) Jack might got unexpected control result in his first and fifth because of the following reasons:

i) In the first case he might have mistakenly chosen plant sequence primer instead of GMO primer.

ii) Since in the fifth case the template is GMO positive PCR should work either in GMO or plant primers. He got negative result which means either PCR master mix composition has problem or there in problem in PCR program which he set.

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