PCR Lab:
For this project we took our own DNA from our cheek cells and went through a number of steps to get the data that shows when your DNA is put into gel red for 72 hours, it will react and a line will form to show you weather your DNA +/+, -/-, or +/-. This will then lead to you finding where your DNA is from.
Purpose: The purpose of this lab was to get lab experience, see the history of our DNA, and learn about lab practices common in biotechnology today.
Hypothesis: I hypothesized I would not get results because the percentage rate is very low. The dependent variable was the DNA because no one has the same DNA. The independent variable is everything else, human error, and all the lab procedures. In this experiment we were testing to see if our DNA results showed as -/-, +/+, or +/-. IN PACKET
Procedure: We followed the procedures given with Babec Alu PCR 2017.
Data and Observation: I predicted I would not get results and I was right. Only four people in our class got back results, two people with -/-, one person with +/+, and one person with +/-. If you got results from your DNA, it can show you your ancestors and where you came from.
During the lab I observed that it took a very steady hand and concentration to put your DNA into the wells with the micropipette. Was difficult to pour original DNA fluid, but having the leave the clump during the beginning of the lab. ADD MORE
Analysis and Discussion: Data shows that when the DNA is put into gel red for 72 hours, it will react and a line will form to show you weather your DNA is +/+, +/-,, or -/-. This will then lead to you finding where your DNA is from.
Some errors that occurred were not spinning the DNA in the microcentrifuge long enough, or the number of tubes being uneven while spinning them. This could result in the cells not splitting up enough leading to not having enough DNA. Making sure to put the right amount of dye into the very small tube. Keeping everything cold in ice and not spilling any components needed. Making sure not to break the gel while using pipet to insert DNA and leaving DNA in heat block at 99 degrees celsius for 10 minutes, not a minute more or less. Putting in the right amount of master and primer mix into your DNA tubes and knowing where you inserted your DNA into the gel.
Steps: 2% agarose gel ran at 150 u for 20min and stained using gel red for 72 hours. Lane 1a and 2a have 100 bp ladder. Lane 1 B-H and 2 D-H have 20 ml of a 50 ml DNA / 10 ml loading dye solution. Lane 1H had sample I worked with
Hypothesis: I hypothesized I would not get results because the percentage rate is very low. The dependent variable was the DNA because no one has the same DNA. The independent variable is everything else, human error, and all the lab procedures. In this experiment we were testing to see if our DNA results showed as -/-, +/+, or +/-. IN PACKET
Procedure: We followed the procedures given with Babec Alu PCR 2017.
Data and Observation: I predicted I would not get results and I was right. Only four people in our class got back results, two people with -/-, one person with +/+, and one person with +/-. If you got results from your DNA, it can show you your ancestors and where you came from.
During the lab I observed that it took a very steady hand and concentration to put your DNA into the wells with the micropipette. Was difficult to pour original DNA fluid, but having the leave the clump during the beginning of the lab. ADD MORE
Analysis and Discussion: Data shows that when the DNA is put into gel red for 72 hours, it will react and a line will form to show you weather your DNA is +/+, +/-,, or -/-. This will then lead to you finding where your DNA is from.
Some errors that occurred were not spinning the DNA in the microcentrifuge long enough, or the number of tubes being uneven while spinning them. This could result in the cells not splitting up enough leading to not having enough DNA. Making sure to put the right amount of dye into the very small tube. Keeping everything cold in ice and not spilling any components needed. Making sure not to break the gel while using pipet to insert DNA and leaving DNA in heat block at 99 degrees celsius for 10 minutes, not a minute more or less. Putting in the right amount of master and primer mix into your DNA tubes and knowing where you inserted your DNA into the gel.
Steps: 2% agarose gel ran at 150 u for 20min and stained using gel red for 72 hours. Lane 1a and 2a have 100 bp ladder. Lane 1 B-H and 2 D-H have 20 ml of a 50 ml DNA / 10 ml loading dye solution. Lane 1H had sample I worked with
Content:
DNA: a molecule that carries the genetic instructions used in the growth, development, functioning and reproduction of all known living organisms
DNA Ladder: a set of standards that are used to identify the approximate size of a molecule run on a gel
Alu Repeat: Alu repeats are approximately 300 base pairs in length. They got their name from the fact that most carry within them the base sequence AGCT which is the recognition site for the Alu I restriction endonuclease, a type of enzyme that cuts DNA at a specific site. There are over 500,000 Alu repeats scattered throughout the human genome. On average, one can be found every 4,000 base pairs along a human DNA molecule." BABEC ALU PCR 2017 Lab
Gel Electrophoresis: a technique used in laboratories in order to separate macromolecules based on size, applies a negative charge so proteins move towards a positive charge. For DNA, smaller sequences move faster because they contain less proteins
PCR: Polymerase chain reaction is a technique to amplify a single copy or a few copies of a segment of DNA, creating millions of copies of a particular DNA sequence
Allele: a gene form from mutation
DNA: a molecule that carries the genetic instructions used in the growth, development, functioning and reproduction of all known living organisms
DNA Ladder: a set of standards that are used to identify the approximate size of a molecule run on a gel
Alu Repeat: Alu repeats are approximately 300 base pairs in length. They got their name from the fact that most carry within them the base sequence AGCT which is the recognition site for the Alu I restriction endonuclease, a type of enzyme that cuts DNA at a specific site. There are over 500,000 Alu repeats scattered throughout the human genome. On average, one can be found every 4,000 base pairs along a human DNA molecule." BABEC ALU PCR 2017 Lab
Gel Electrophoresis: a technique used in laboratories in order to separate macromolecules based on size, applies a negative charge so proteins move towards a positive charge. For DNA, smaller sequences move faster because they contain less proteins
PCR: Polymerase chain reaction is a technique to amplify a single copy or a few copies of a segment of DNA, creating millions of copies of a particular DNA sequence
Allele: a gene form from mutation
A B C D E F G H
A B C D E F G H
Reflections:
I think during this project I improved on working a lot more and being more efficient with my time. I learned about myself on how I work better doing my own task or something separate from the group, because your group was half of the class. A new skill I gained during this project was how to be more efficient with my time and work more by myself and being more focused. For example when most people in the group were inserting their DNA into the gel solution, I was putting the food coloring into my DNA, taking my time so I would make any errors. I think I could have improved on my listening skills. For example, I sometimes wouldn't listen or read the instructions thoroughly, so someone in my group would have to describe to me what we were doing again. Overall, I think the project went well and helped me improve on my work ethic skills and it was a fun experience.
I think during this project I improved on working a lot more and being more efficient with my time. I learned about myself on how I work better doing my own task or something separate from the group, because your group was half of the class. A new skill I gained during this project was how to be more efficient with my time and work more by myself and being more focused. For example when most people in the group were inserting their DNA into the gel solution, I was putting the food coloring into my DNA, taking my time so I would make any errors. I think I could have improved on my listening skills. For example, I sometimes wouldn't listen or read the instructions thoroughly, so someone in my group would have to describe to me what we were doing again. Overall, I think the project went well and helped me improve on my work ethic skills and it was a fun experience.