Gel Electrophoresis Lab- Dominic Lal

Video
This experiment uses micropipettes, agarose gel, DNA samples, and the gel electrophoresis chamber. This video discusses the purpose behind gel electrophoresis as well as the experimental set up. It also reviews the use of calculations to determine the base-pair lengths when given the distance traveled. Overall this experiment is useful for forensic scientist when comparing DNA between various people.

Example Data
Below is an example gel that has gone through the gel electrophoresis experiment. The chart contains the Base-pair of DNA that was cut by restriction ezyme Hind III. To find the distance traveled, a ruler could be used to measure the distance traveled from the well. The completed Hind III chart is expressed as Figure 3. After the information of Hind III was created, a scatter plot graph was used to make the equation needed for finding the BP length. This will be further explained in the sample calculation section.





Hind III

Figure 3
This scatter plot was compiled from the data given above where the base-pair length was the y-axis and the distance traveled in cm was the x-axis. The line of degression is shown as the dotted line and this expresses the formula y= -3367.2x+24973. So the x may be plugged in to find the y. This equation was used to complete the BamHI and EcoRi sections of the table.

BamHI                                                             EcoRI

Example Calculations
This is the completed chart which was found by measuring the distance (in cm) that the DNA strands moved. After the distance was compiled, the distance(x) was plugged into the formula found by the scatter plot above to find the BP

length(y). So from the data that was found above a scatter plot was created in order to find the y=mx+b formula that can be used to find the base-pairs of other strands after finding the distance traveled. In this particular example the formula is y=-3367.2x+24973.

Sample Problem: So if the distance traveled of a BamHI is 4.2 and the equation that Hind III information gives is y=-3367.2x+24973 what is the Base-pair length?

So 4.2 must be plugged into the equation as x to calculate to 10,494.04

Practice Problem 1: So if the distance traveled of a EcoRI is 2.8 and the equation that Hind III information gives is y=-3367.2x+24973 what is the Base-pair length?

Practice Problem 2: So if the distance traveled of a BamHI is 4.7 and the equation that Hind III information gives is y=-3367.2x+24973 what is the Base-pair length?

Quizlet
https://quizlet.com/82152429/gel-electrophoresis-lab-study-guide-flash-cards/

Restriction enzymes
Restriction enzymes are needed to cut the DNA sequences. And each restriction enzyme cuts at different points. An enzyme from bacteria that can recognize specific base sequences in DNA and cut the DNA at that site (the restriction site). A restriction enzyme acts as a biochemical scissors. Also called a restriction endonuclease.

Evolution
The gel electrophoresis is a process where the differences in the strands of DNA shows the diversity. Over time, the common ancestor evolved to have different sequences of DNA. So evolution changed populations of organisms over time due to environmental factors.

Amplification of DNA
The PCR or polymerase chain reaction can produce copies of a specific target segment of DNA. It goes through a three-step cycle where it is first heated, then cooled, and then replicated. When this occurs it creates a chain reaction that produces an exponentially growing population of identical DNA molecules. This is similar to gel electrophoresis because it is used to create more DNA, which can be analyzed through gel electrophoresis. If there is not enough of the DNA sample found to go into the gel electrophoresis procedure, the DNA could be amplified to produce more so that the DNA can be tested and anaylzed.

Practice Problem 1: 15,544.84

Practice Problem 2: 9,147.16