Based on the lab experiement for DNA extraction of Human Buccal Cells with the i
ID: 166339 • Letter: B
Question
Based on the lab experiement for DNA extraction of Human Buccal Cells with the intention of sequencing our own DNA , a total of 2 Electrophoresis Gels Runswhere completed. Based on the PCR A & B Gel electrophoresis results given, Please provide short, concise, and detailed explanation in respect to the movement of DNA through the agarose gel (1.5 %)( i.e. speed/ size/ etc. of fragments, ~size of Base pairs), what the results visualized represent, some possible reasons/sources of error to support & explain why only PCR product A bands { except for Student #6 who displayed both A & B in Bottom row for Ladders 8 & 9, WHY??} were observed during Gel Run #1 where on the contrary, Both PCR products A & B bands were displayed & observed in both rows & ladders for ALL students in Gel Run #2.
FYI ** PCR was carried out twice for all the students and then loaded in the two gels **
Urgent: Best answer gets rated/need Asap** ty!
GEL RUN #1: Red color observed in Row 2 of Ladder #9 (student #6) is due to Artifact right? LMK
GEL ELECTROPHORESIS RUN #1 FOR PCR PRODUCTS (ONLY PCR PRODUCT A VISIBLE IN LADDERS except for Student #6 which displayed both Products A & B)
**I have provided loading orders for reference**
Loading Order:
Lane (top row)
1
ladder
2
Student #1
A
3
Student #1
B
4
5
Student #2
A
6
Student #2
B
7
8
Student #3
A
9
Student #3
B
10
Bottom row
1
Ladder
2
Student #4
A
3
Student #4
B
4
5
Student #5("ME!")
A
6
Student #5("ME!")
B
7
8
Student #6
A
9
Student #6
B
10
.
Gel RUN #2:
GEL ELECTROPHORESIS RUN #2 FOR PCR PRODUCTS (BOTH PCR PRODUCTS A & B VISIBLE IN LADDERS RESULTING IN A MORE SUCCESSFUL GEL RUN**)
**I have provided loading orders for reference**
Loading Order
Lane (top row)
1
ladder
2
Student #1("Me!")
A
3
Student #1("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
Loading Order:
Lane (top row)
1
ladder
2
Student #1
A
3
Student #1
B
4
5
Student #2
A
6
Student #2
B
7
8
Student #3
A
9
Student #3
B
10
Explanation / Answer
In Gel Run #1, the PCR products might not have amplified in case of all students, that might be the reason why there are missing bands.
Red color observed in Row 2 of Ladder #9 (student #6) is not an artifact its a PCR product.
Gel Run #1 is run at a high voltage and there is more ethidium bromide concentration in the gel. Some of the DNA in Gel Run#1 is still stuck in the wells it not run properly at all.
The size of the fragments cannot be determined as the ladder details are not given.