Procedure:
Purpose of Lab 4A: To make 10 milliliters of 5 M NaCl solution. To make 100 mL of TE buffer: 10 mM TRIS, 1mM EDTA (DNA storage solution)
Purpose of Lab 4B: Can DNA be spooled out of solution? What does DNA look like? What are some of its unique properties? What yield of DNA can be recovered during the isolation?
Purpose of Lab 4I: To prepare and pour an agarose gel for DNA fragment analysis.
Purpose of Lab 4J: What was the appearance of different DNA samples on an agarose gel?
Materials:
nalytical Balance
Tabletop Balance
Weigh Paper/Boat
Scoops
Sodium Chloride
Capped Tubes(15ml) and Rack
TRIS
EDTA
Sodium Hydrochloric Acid
Sodium Hydroxide
Granulated Cylinder(100ml)
pH strips
50ml Beakers
DNA(Salmon Testes)
2ml Pipet and Pump
Micropipet, P-1000, and Tips
95% Ethanol
Glass Rods
TAE buffer, 40X
600ml Beakers
Agarose
250ml bottle
Microwave Oven
Hot Hands Protector
Safety Glasses
Gel Box
Gel Tank
Power Supply
Gloves
Ethidium Bromide
Microcentrifuge
6x Loading Dye
Lab 4A:
Part I: Preparing 5M of NaCL
1. Measure out 2.92 g of NaCl and add to tube
2. QS water to (add water until you reach) 10 mL
3. Label for later use
Part II: Preparing TE Buffer
1. Add 0.1576 g of TRIS and 0.037224 g of EDTA to a beaker
2. Add 80 mL of deionized water to solution
3. Add HCl to raise pH or NaOH to lower pH until solution has a pH between 7.5 and 8.5
4. QS water to 100 mL
5. Label for later use
Lab 4B:
1. Add 1 mL DNA and 1 mL TE into a beaker.
2. Add 5M NaCl
3. Add 4 mL ETOH by pouring down side (forms two layers)
4. Using a glass rod, spool DNA
5. Place DNA in a tube with 2 mL TE
Lab 4I:
1. Add 0.4g agarose powder and QS TAE to 100 mL
2. Heat solution until agarose is dissolved.
3. Let solution cool and then pour into gel box (with combs)
Lab 4J:
1. Submerge gel and electrodes in TAE
2. Add 20 uL DNA and 4 mL of 6x loading dye into a tube
3. Centrifuge solution
4. Load solution into gel
5. Apply charge to gel for approximately 45 minutes
6. Stain with Ethidium Bromide, rinse, and observe
Data Analysis:
This lab did not work. After the staining with Ethidium Bromide, no DNA showed up in the gel. This happened to all of the groups. This happened due to our dye expiring. To retest we would need need to make a whole new batch of dye.
Conclusion:
In this lab, our group tried to separate DNA from a solution. Seeing the DNA separate reinforces our knowledge about the properties of DNA. These methods can be applicable to many biotech and forensic lab studies.
Reflection:
I believe that our group worked pretty well together. However, some of the work was divided up inefficiently. Some people would be working while the rest of the group was just sitting there. Instead, we could have divided the tasks evenly among the group before we began the experiments. I liked this lab over all and enjoyed working with my group.
Purpose of Lab 4B: Can DNA be spooled out of solution? What does DNA look like? What are some of its unique properties? What yield of DNA can be recovered during the isolation?
Purpose of Lab 4I: To prepare and pour an agarose gel for DNA fragment analysis.
Purpose of Lab 4J: What was the appearance of different DNA samples on an agarose gel?
Materials:
nalytical Balance
Tabletop Balance
Weigh Paper/Boat
Scoops
Sodium Chloride
Capped Tubes(15ml) and Rack
TRIS
EDTA
Sodium Hydrochloric Acid
Sodium Hydroxide
Granulated Cylinder(100ml)
pH strips
50ml Beakers
DNA(Salmon Testes)
2ml Pipet and Pump
Micropipet, P-1000, and Tips
95% Ethanol
Glass Rods
TAE buffer, 40X
600ml Beakers
Agarose
250ml bottle
Microwave Oven
Hot Hands Protector
Safety Glasses
Gel Box
Gel Tank
Power Supply
Gloves
Ethidium Bromide
Microcentrifuge
6x Loading Dye
Lab 4A:
Part I: Preparing 5M of NaCL
1. Measure out 2.92 g of NaCl and add to tube
2. QS water to (add water until you reach) 10 mL
3. Label for later use
Part II: Preparing TE Buffer
1. Add 0.1576 g of TRIS and 0.037224 g of EDTA to a beaker
2. Add 80 mL of deionized water to solution
3. Add HCl to raise pH or NaOH to lower pH until solution has a pH between 7.5 and 8.5
4. QS water to 100 mL
5. Label for later use
Lab 4B:
1. Add 1 mL DNA and 1 mL TE into a beaker.
2. Add 5M NaCl
3. Add 4 mL ETOH by pouring down side (forms two layers)
4. Using a glass rod, spool DNA
5. Place DNA in a tube with 2 mL TE
Lab 4I:
1. Add 0.4g agarose powder and QS TAE to 100 mL
2. Heat solution until agarose is dissolved.
3. Let solution cool and then pour into gel box (with combs)
Lab 4J:
1. Submerge gel and electrodes in TAE
2. Add 20 uL DNA and 4 mL of 6x loading dye into a tube
3. Centrifuge solution
4. Load solution into gel
5. Apply charge to gel for approximately 45 minutes
6. Stain with Ethidium Bromide, rinse, and observe
Data Analysis:
This lab did not work. After the staining with Ethidium Bromide, no DNA showed up in the gel. This happened to all of the groups. This happened due to our dye expiring. To retest we would need need to make a whole new batch of dye.
Conclusion:
In this lab, our group tried to separate DNA from a solution. Seeing the DNA separate reinforces our knowledge about the properties of DNA. These methods can be applicable to many biotech and forensic lab studies.
Reflection:
I believe that our group worked pretty well together. However, some of the work was divided up inefficiently. Some people would be working while the rest of the group was just sitting there. Instead, we could have divided the tasks evenly among the group before we began the experiments. I liked this lab over all and enjoyed working with my group.