DNA Biology and Technology Lab Report
Microbiology: BIO 299
Introduction
Biotechnology is considered to be the science of using living systems and substances derived from them, most commonly referred to as DNA, to produce applications that benefit mankind. During this lab you will learn about both DNA isolation along with gel electrophoresis and how they relate to biotechnology. As more research is done, the subject of biotechnology grows. We can see the massive feats overcome when looking at things like forensics and parental testing. This lab teaches how both Isolation of DNA and Gel Electrophoresis are performed and give you an idea of how they can be useful in the real world.
Materials and Methods
When performing DNA isolation, you will need the following:
· Meat tenderizer
· Sodium chloride
· Water solution
· Strawberry filtrate
· Pipette
· Test Tube
· Glass Rod
· Bucket of Ice
· Ethanol
· Graduated Cylinder
Start by placing your strawberry filtrate and your ethanol into the bucket of ice to ensure it stays cold. Then place your test tube into the ice and transfer some of the strawberry filtrate into the tube. In the graduated cylinder combine the meat tenderizer and water. Transfer a few mL into the test tube and let it react for 10 minutes. Once this time has passed, add 95% ethanol into the test tube. This should case the DNA to precipitate. Now, turning in a clockwise motion, spool the DNA precipitate onto the glass rod. When removed you should see a stringy, gelatinous material on the glass rod. This indicates this part of the lab was done correctly.
In order to perform the second section of this lab, gel electrophoresis, you will need:
· Hot Liquid Agar
· A power supply
· Electrophoresis buffer
· Ethidium Bromide
· Micro Pipette
· DNA Samples
· Masking Tape
· Gel Comb
· Gel Casting Tray
· Electrophoresis Chamber
· Pipette Microtips
To begin the second half of this lab you must first assemble the gel tray. Start by placing masking tape on both ends and adding the comb to create wells for the DNA you will add later. Next, add Ethidium Bromide to the hot liquid agar and gentle pour into the gel tray. Wait a minimum of 10 minutes in order to let the gel cool before moving forward. Once the gel has solidified, carefully remove the tape and comb.
Next you must prepare the electrophoresis chamber and load the DNA samples. To begin this start by placing the gel into the electrophoresis chamber and fill the chamber with the electrophoresis buffer. Add your DNA samples into the gel using the pipette, each of them going into their own well. Be sure to use a new pipette tip with each sample to avoid cross-contamination. Place your control samples (TNF1 and TNF2) into their own well also. Cover your electrophoresis chamber and turn on the power source and set the voltage to 70V. Press play and let run into the DNA bands have sufficiently separated. Feel free to check as time passes. Once the gel has ran for the necessary time, plate under a UV table and turn on the light. You then must identify the genotypes of the three test samples by matching them with the controls.
Results
When performing the DNA isolation correctly we are able to collect a sample using a technique called spooling. Temperature is a very important part of this lab and you will not be able to see results if everything is not kept cold. The coolness allows for stabilization of the hydrogen bonds.
When completing the gel electrophoresis, you are able to see which DNA sample matches with which control sample. It is also possible to see that one of the DNA samples is a combination of the two controls. The electrophoresis chamber properly separated the DNA.
Conclusion
This lab allowed for a better understanding of how DNA works and how it can be used in the real world. We separate DNA by way of DNA isolation. This allows for a pure DNA sample to be retrieved. By using gel electrophoresis, we are able to determine if DNA is similar or related. Gel Electrophoresis isolates DNA based on the size. Both methods are regularly used for forensics and are quite helpful in solving crimes.
