Lab 4
Purpose:
a. Make 10 ml of 5M NaCl solution and to make 100 ml of TE buffer (10mM TRIS and 1mM EDTA)
b. Investigate whether or not DNA can be spooled. Figure out it's appearance, its properties, and what yield of DNA can be found
i. Create and prepare an agarose gel for DNA fragment analysis
j. Find appearance differences on separate DNA samples and effects they have on the agarose gel
a. Make 10 ml of 5M NaCl solution and to make 100 ml of TE buffer (10mM TRIS and 1mM EDTA)
b. Investigate whether or not DNA can be spooled. Figure out it's appearance, its properties, and what yield of DNA can be found
i. Create and prepare an agarose gel for DNA fragment analysis
j. Find appearance differences on separate DNA samples and effects they have on the agarose gel
Calculations: (Molarity) x (Volume) x (Formula Weight) = grams of substance needed
NaCl- (5 M)(0.01 L)(58.44 g/mol) = 2.92 grams TRIS- (0.01 M)(0.1 L)(157.6 g/mol) = 0.158 grams EDTA- (0.001 M)(0.1 L)(372.24 g/mol) = 0.037 grams |
Conversions to Liters and Moles:
|
Materials:
Analytical Balance Tabletop Balance Weigh Paper/Boat Scoops Sodium Chloride Capped Tubes(15ml) TRIS EDTA Sodium Hydrochloric Acid Sodium Hydroxide Granulated Cylinder(100ml) |
Safety Glasses
Gel Box Gel Tank Power Supply Gloves Ethidium Bromide Microcentrifuge 6x Loading Dye Distilled Water 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 Tube Rack |
Procedure:
First step measure out 2.92 g of sodium and put it into 15mL of deionized water then vortex it until no more salt seen. Next weigh out 0.37 of EDTA and 1.58 Tris wi and 80mL water into a test tube to make 100mL TE. Measure PH make PH 8 using sodium hydroxide then put 1mL Te buffer with salmon sperm sample keep chilled. Add 5 micro liters then add 5mL of Ethanol spool. Put DNA in test tube. Then make 500mL from TAE buffer from 40x concentrate. Weigh 0.48 agrose and put in flask dissolve. Put into mold and let it cool(TAE covers it then stain DNA and inject into the mold and send voltage through.
Data Analysis - From our data analysis the first time we ran the test the results were negative so then we had to run the test again with our solution fixed. The DNA moved through the gel and looked like small oval like bands.
First step measure out 2.92 g of sodium and put it into 15mL of deionized water then vortex it until no more salt seen. Next weigh out 0.37 of EDTA and 1.58 Tris wi and 80mL water into a test tube to make 100mL TE. Measure PH make PH 8 using sodium hydroxide then put 1mL Te buffer with salmon sperm sample keep chilled. Add 5 micro liters then add 5mL of Ethanol spool. Put DNA in test tube. Then make 500mL from TAE buffer from 40x concentrate. Weigh 0.48 agrose and put in flask dissolve. Put into mold and let it cool(TAE covers it then stain DNA and inject into the mold and send voltage through.
Data Analysis - From our data analysis the first time we ran the test the results were negative so then we had to run the test again with our solution fixed. The DNA moved through the gel and looked like small oval like bands.
Conclusion
The group I was in this lab was not a group I would not normally work in. I worked with Bergo, Cantor, and Maga.
in this group we got alot of things done fast and efficiently. We may of messed up a couple times but we got over our obstacles. One thing we could've done better is people getting different materials rather than all of us waiting at the same station. Another would be more communication because sometimes we will be not sure what someone will be doing.
The group I was in this lab was not a group I would not normally work in. I worked with Bergo, Cantor, and Maga.
in this group we got alot of things done fast and efficiently. We may of messed up a couple times but we got over our obstacles. One thing we could've done better is people getting different materials rather than all of us waiting at the same station. Another would be more communication because sometimes we will be not sure what someone will be doing.