Embedded Files
Purpose:
The purpose of the pGlo Bacterial Transformation Lab is to learn how to insert a gene into bacteria and figure out whether the bacteria took on the gene. This has a variety of uses in our future learning both safe handling of bacteria and bacteria cultivation, not to mention getting bacteria to accept a gene that we want it too. This improves our problem solving skills and logical reasoning to create reasonable conclusions.
Materials:
Agarose, 0.48 g
Graduated cylinder, 100-mL
Biological dye solutions, 0.5 mL,
Glycerin, 3 mL
Microwave or stirring hot plate
Tris-acetate electrophoresis buffer, 50X, 10 mL
Water, deionized or distilled (DI)
Pipets, graduated, disposable, 7
Balance, 0.01-g precision
Pipets, needle-tip, disposable or micropipets with tips, 6
Cotton, non-absorbent or foam plug
Power supply
Electrophoresis chamber
Spot plate or reaction plate
Erlenmeyer flask, 500-mL
Stirring rod
Erlenmeyer flask, borosilicate, 125-mL
Thermometer, 0–100 °C
Gel casting tray with well comb
Toothpicks, 6
Graduated cylinder, 25-mL
Weighing dish, small
Observations:
You cant fill the micropipet all the way because it will splat out when you release it
You have to be careful when you make your gel and place it because it's fragile
You have to go slow when releasing the dye because it will get everywhere
Don't push the micropiet too deep because it will go into the gel and have invalid results
Analysis:
Shows gel prepared to get measured
Shows the process of how to measure
Measurements and names of dye
Conclusion:
Conclusion:
In this lab I utilized my understanding of biology and chemistry techniques to perform the lab quickly and efficiently. After doing measurements for each dye in the lab and comparing my results I say with confidence that Unknown 1 is made of Bromothymol Blue, Methyl Red, and Thymol Blue. According to my data Unknown two is made of Malachite Green, Safranin O and Bromothymol Blue. I know these are correct because they had about the right distance and the right charge.
Reflection:
Reflection:
This lab was really fun! It was quick and painless, the only difficulty I ran into was putting the dyes into the gel, sometimes I would mess it up and the dye would end up mostly in the solution rather than the gel. I wish we had one or two more days to run more tests so I could get an average for the dyes rather than just one test because human error in extremely present in many ways like putting too much dye in, putting it in the wrong spot, so many tiny things that could have been messed up. But I really did have fun with this lab and I cant wait for more techniques in the coming weeks like this one because this was fun!
Lab Technique's:
IMG_9046.MOVThis technique is Dye Electrophoresis and it works by running electricity through a medium and because the dye is in the middle ina gel it moves towards a charge at a certain speed based on the dyes properties. You can then use that distance in X time to compare other results and identify what a composite dye is made out of using cross referencing.
IMG_9165.MOVThe title of this lab and the technique—Micropipetting— this technique is very simple yet tricky to get right. You need to put the micropipet onto a disposable tip and insert the tip into the dye and press the top not all the way but to where it stops, and then very carefully put the tip into the well and push down onto the top all the way. Then put the tip into the garbage.
IMG_9160.movLab Sheets:
Lab Sheets:
Dyeing for Electrophoresis.PDFLab sheet for the experiment
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