Please help. The deadline is in about 3 hours. (If I submit an answer, I don\'t
ID: 174072 • Letter: P
Question
Please help. The deadline is in about 3 hours. (If I submit an answer, I don't know if it's right or wrong until after the deadline.)
Use the following figure to answer the next 3 questions. P3 P1 cat P2 P4 lacz Using the figure above when cloning the cat gene into pBLU, you want to identify the orientation in which cat was inserted. Using a PCR reaction, you use the primer combinations of 1 and 2, 1 and 3, and 1 and 4. You see that there is an expected product size using primers 1 and 3, no product from 1 and 2, and a product from 1 and 4. How did cat insert? In the opposite orientation as lacz O In the same orientation as lacz Nothing was inserted Multiple copies were inserted Submit Answer Tries 0/99 Using the figure above, you clone the cat gene into pBLU. To determine the manner in which cat was inserted, you use the primer combinations 1 and 2, 1 and 3, and 2 and 3. You see a product from every reaction, what happened? Cat inserted in the opposite orientation as lacz O Multiple copies of cat were inserted cat inserted in the same orientation as lacz O Nothing was inserted Submit Answer Tries 0/99 Using the figure above when cloning the cat gene 1 and 2 and no product identify and orientation in which cat was inserted. Using a PCR reaction, 4. How did cat insert? of 1 and 2, 1 combinations and primer and the 3, and use 1 You you 4. see no product from expected from 1 and into product than pBLU, you a smaller want to from 1 see the Nothing was inserted In the opposite orientation as lacz In the same orientation as lacz O Multiple copies were inserted Submi Answer Tries 0/99Explanation / Answer
1) answer: 1st option.
the product from 1 & 4confirms the presence of the insert within the lac gene. 1& 3 both are forward primers . 2 primers priming the same orientation will never amplify the gene. But 1&3 has shown a product amplification. this indicates a change in orientation of the gene.
2)Answer is the 2nd option.
1&2 will give a product when the cat gene is inserted in the same orientation as that of lacZ, and the 2&3 combination will give product irrespective of the orientation of insertion. But 1&3, giving product means , the orientation is changed. therefore, there are multiple copies in both orientations are inserted.
3) The answer is the 1st option.
no product from 1&3 and 1&2 means an absence of cat gene in both orientations. a smaller product from 1&4 means only the lac gene sequence in between the primers are amplified. thus confirming the absence of insert.