A total of 2 Electrophoresis Gels Runs where completed based on the lab experime
ID: 166432 • Letter: A
Question
A total of 2 Electrophoresis Gels Runs where completed based on the lab experiment: DNA extraction of Human Buccal Cells with the intention of sequencing our own DNA. Based on the PCR A & B Gel electrophoresis results of six different students which are given, please provide a short, concise, and detailed explanation in respect to the movement of DNA through the agarose gel (1.5 %)( e.g. speed, size of fragments, ~size of Base pairs, etc.). Please dont forget to size all bands other than template ladder ty!
What exactly do the results visualized represent? For both Gel Run # 1 & # 2.
Regarding Gel run # 1; what are some of the possible reasons including sources of error that may support & explain why only PCR product A was observed in all ladders of both the top & bottom row with the exception of Student # 6 whose PCR A & B bands were present & seen @ ladder 8 & 9(bottom row) whereas all other students only displayed PCR Product A in this particular run.
Unlike Gel run # 1, When a second gel was run by our professor, Both PCR products A & B bands were displayed & observed in both top & bottom rows & all ladders by all students results in Gel Run #2. Why ??
Urgent: Best answer gets rated/need Asap** ty!
GEL RUN #1: (Red color observed in bottom Row of Ladder #9 (student #6) is due to Artifact !!)
[Table 1] Loading Order
Lane (top row)
1
ladder
2
Alyssa
A
3
Alyssa
B
4
5
Brittani
A
6
Brittani
B
7
8
Hunter
A
9
Hunter
B
10
Bottom row
1
Ladder
2
Chris
A
3
Chris
B
4
5
CR (ME!)
A
6
CR (ME!)
B
7
8
Raj
A
9
Raj
B
10
Gel Run # 2:
** BOTH PCR PRODUCTS A & B VISIBLE IN LADDERS RESULTING IN A MORE SUCCESSFUL GEL RUN**
GEL RUN #2: Red color observed in bottom Row of for Ladders (student #6) is due to Artifact !
Loading Order for reference
[Table 1] Loading Order
Lane (top row)
1
ladder
2
CR (Me!)
A
3
CR (Me!)
B
4
Student #2
A
5
Student #2
B
6
Student #3
A
7
Student #3
B
Bottom row
1
ladder
2
Student #4
A
3
Student #4
B
4
Student #5
A
5
Student #5
B
6
Student #6
A
7
Student #6
B
[Table 1] Loading Order
Lane (top row)
1
ladder
2
Alyssa
A
3
Alyssa
B
4
5
Brittani
A
6
Brittani
B
7
8
Hunter
A
9
Hunter
B
10
Bottom row
1
Ladder
2
Chris
A
3
Chris
B
4
5
CR (ME!)
A
6
CR (ME!)
B
7
8
Raj
A
9
Raj
B
10
Gel Run # 2:
Explanation / Answer
Gel Run#1 and Gel Run #2 clearly represented the separation of DNA fragments based on their size. The size will be calculated from the ladder separation in agarose gel electrophoresis. Smaller fragments will move faster than larger fragments. Here, most of the fragments have high molecular weight except few ones. Ethidium bromide acts as a visualizing agent and we can easily visualize the DNA under UV light. PCR reaction mainly used to amplification of your target genes. Here, the PCR product observed in all top and bottom row except in student #6 and this indicated that your target genes are well amplified through PCR reaction. Whereas in student #6 failed to exhibit the result due to improper handling or adding more amount of primers,dNTPS etc.
Related Questions
drjack9650@gmail.com
Navigate
Integrity-first tutoring: explanations and feedback only — we do not complete graded work. Learn more.