DNA Labs (4a, 4b, 4i, 4j)
Purpose:
4a: To prepare the NaCl solution and to make the TE buffer for the next labs.
4b: To spool DNA and put it in the TE buffer.
4i: To prepare and pour an agarose gel for DNA fragment analysis.
4j: To find what different DNA samples look like in agarose gel.
Materials:
4a: To prepare the NaCl solution and to make the TE buffer for the next labs.
4b: To spool DNA and put it in the TE buffer.
4i: To prepare and pour an agarose gel for DNA fragment analysis.
4j: To find what different DNA samples look like in agarose gel.
Materials:
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Procedure:
Lab 4a:
Making the NaCl:
Making the TE buffer:
Lab 4b:
Lab 4i:
Lab 4j:
Data Analysis:
This is the first class project that didn't work. The DNA didn't show up after we stained with the ethidium bromide. Everyone had this problem even 2/3 period, so we brainstormed some ideas the next day to see if we could salvage this experiment. The most likely reason was that the dye was expired. So Dr. LB made a new dye and did the last step again. Hopefully the experiment will work this time.
Conclusion:
We tried to separate DNA from a solution but failed. We learned that projects may go wrong and what to do when that happens. We also got to see actual DNA. This is a technique used by forensic scientists to determine the culprit of a crime.
Reflection:
This was a great project because we got to have bigger groups. Also it was over the course of a couple of days. But there were some problems. One problem was that some people did most of the work while some did barely did anything. Another problem was that some of the ingredients didn't work as we could see in the last part of the experiment. Lastly, the group kept on screwing around so it took a while for this to get done. So this was an OK project.
Lab 4a:
Making the NaCl:
- Measure 2.92 grams of NaCl and put it in tube
- Add water until you reach 10 mL
- Label the tube
Making the TE buffer:
- Measure 0.1576 grams of TRIS and 0.037 grams EDTA to a beaker
- Add 80 mL of deionized water to the solution
- Add either HCl or NaOH to raise or lower the pH level until it is between 7.5-8.5
- Add water until you reach 100 mL
- Label
Lab 4b:
- Add 1 mL DNA and 1 mL TE into a beaker
- Add 5 M of NaCl
- Add 4 mL of ETOH to form 2 layers
- Spool DNA using glass rod
- Place DNA in tube with 2 mL TE buffer
Lab 4i:
- Add 0.4 grams agarose powder and add TAE until it reaches 100 mL
- Heat solution until dissolved
- Let solution cool and put in gel box with combs
Lab 4j:
- Submerge gel and electrodes with TAE
- Add 20 uL DNA and 4 mL of 6x loading dye into tube
- Centrifuge solution
- Load solution into gel
- apply charge to gel and wait for about 45 minutes
- Stain with Ethidium Bromide, rinse and observe
Data Analysis:
This is the first class project that didn't work. The DNA didn't show up after we stained with the ethidium bromide. Everyone had this problem even 2/3 period, so we brainstormed some ideas the next day to see if we could salvage this experiment. The most likely reason was that the dye was expired. So Dr. LB made a new dye and did the last step again. Hopefully the experiment will work this time.
Conclusion:
We tried to separate DNA from a solution but failed. We learned that projects may go wrong and what to do when that happens. We also got to see actual DNA. This is a technique used by forensic scientists to determine the culprit of a crime.
Reflection:
This was a great project because we got to have bigger groups. Also it was over the course of a couple of days. But there were some problems. One problem was that some people did most of the work while some did barely did anything. Another problem was that some of the ingredients didn't work as we could see in the last part of the experiment. Lastly, the group kept on screwing around so it took a while for this to get done. So this was an OK project.