DNA Lab Write Up 10-11-14
Purpose-
To make:
10mL 5M NaCL, then 100mL TE 10mM Tris, 1mM EDTA.
Why:
We did this to observe the action of spooling DNA.
To make:
A solution of TAE(Tris-acetate-EDTA) and 8% agarose in 1 x TAE.
Why:
To make a mold for a gel.
To learn:
What is the appearance of different DNA samples on an agarose gel?
Materials-
Our DNA bands did not appear. After a long discussion, we believe we had a fault in our stain. The stain must have deteriorated over the last year due to possible exposure to UV rays, but we can't be sure. We also thought there may be a broken scale factor, even though it wouldn't have effected all the solutions. Our only way to find out is to hand-make a stain to use to test the issue if it was the problem. Other possibilities we thought of was human error, although said before, it wouldn't have effected all the student experiments.
Conclusion-
The importance of isolating DNA from a solution and separating it on a gel to scientists is that they can use the isolated DNA to detect genetic disorders, identify a suspect in forensics with their fingerprint, and use the DNA to help studies, like insulin, hormones, and antibiotics. An extraction of DNA can help a scientist study a specific gene. This gene can help a scientist determine a treatment for the particular disease. As for forensics, DNA can help one identify the suspect in need, most commonly with fingerprints.
Reflection-
My group worked very well. I decided not to be in an ordinary group with my friends but people I knew wouldn't slack during the lab. We all contributed efficiently, having one person doing something at all times until the next initial instructions. The only mistake I made was adding too much water to the pH concentration by accident. This couldn't be fixed unless we redid the whole solution. I can avoid this in the future by knowing more background on the goal of the experiment and being more precise during the adding of substances. My skill at pipetting is a little above average, I could use more help on the micropipettes and what the amounts on the front with the numbers pertain to. I think we could use a little more practice on making solutions and adjusting pH may be on more time would assure our skills.
We eventually, redesigned and successfully did our experiment to see results. The DNA strands were really cool to see with the gel process. I was happy we didn't really do all this work for nothing. Even though we didn't do a whole lot of work it was nice to see some results after a chemical error.
To make:
10mL 5M NaCL, then 100mL TE 10mM Tris, 1mM EDTA.
Why:
We did this to observe the action of spooling DNA.
To make:
A solution of TAE(Tris-acetate-EDTA) and 8% agarose in 1 x TAE.
Why:
To make a mold for a gel.
To learn:
What is the appearance of different DNA samples on an agarose gel?
Materials-
- balance, analytical
- balance, tabletop milligram
- weigh paper
- weigh boat
- lab scoops
- sodium chloride
- tubes
- tube racks
- TRIS
- EDTA
- bottle, 125 mL
- graduated cylinder, 100mL
- pH paper
- hydrochloric acid
- sodium hydroxide
- glass rods
- beakers, 50 mL
- DNA salmon testes
- pipet, 2mL
- pipet pump, blue
- micropipet, p-1000
- micropipet tips for p-1000
- ethanol, 95%
- lab marker pens
- plastic beaker
- TAE buffer concentrate, 40x
- beakers, 600mL
- agarose
- media bottle, 250mL
- microwave oven
- hot hands protector
- gel box
- beakers, 50mL
- water bath, 65 degrees Celsius
- tube rack for 1.7mL tubes
- reaction tubes, 1.7mL
- gel loading dye
- micropipet, p-10
- micropipet, p-100
- microcentrifuge
- power supply
- ethidium bromide
- gel photo imaging system
- paper, thermal
- printer, thermal
- gloves
- glasses, safety, plastic
- weigh the sodium
- put 10mL deionized water into 15mL test tube with 2.92g sodium
- vortex
- weigh EDTA of 0.37g
- weigh Tris for 1.58g
- add Tris and EDTA and 80mL water into a test tube to make 100mL TE
- record pH level
- raise pH to 8 with base
- add sodium hydroxide to adjust pH
- put 1mL TE buffer with salmon sperm sample
- keep everything chilled
- add 500 microliters
- add 5 ml ethanol
- spool and observe DNA
- put DNA into test tube
- make 500mL of 1 x TAE buffer from 40 x concentrate
- weigh 0.4g agarose
- heat flask to dissolve
- prep gel mold
- pour gel into gel mold and let cool
- after cool, remove tape from gel, place in gel tank
- pour TAE over gel until covered; gently remove combs
- prepare samples
- load samples onto gel
- put cover on gel tank, plug into power supply
- run at 110v for 45 minutes
- stain several hours with EtBr; rinse and observe with out light
Our DNA bands did not appear. After a long discussion, we believe we had a fault in our stain. The stain must have deteriorated over the last year due to possible exposure to UV rays, but we can't be sure. We also thought there may be a broken scale factor, even though it wouldn't have effected all the solutions. Our only way to find out is to hand-make a stain to use to test the issue if it was the problem. Other possibilities we thought of was human error, although said before, it wouldn't have effected all the student experiments.
Conclusion-
The importance of isolating DNA from a solution and separating it on a gel to scientists is that they can use the isolated DNA to detect genetic disorders, identify a suspect in forensics with their fingerprint, and use the DNA to help studies, like insulin, hormones, and antibiotics. An extraction of DNA can help a scientist study a specific gene. This gene can help a scientist determine a treatment for the particular disease. As for forensics, DNA can help one identify the suspect in need, most commonly with fingerprints.
Reflection-
My group worked very well. I decided not to be in an ordinary group with my friends but people I knew wouldn't slack during the lab. We all contributed efficiently, having one person doing something at all times until the next initial instructions. The only mistake I made was adding too much water to the pH concentration by accident. This couldn't be fixed unless we redid the whole solution. I can avoid this in the future by knowing more background on the goal of the experiment and being more precise during the adding of substances. My skill at pipetting is a little above average, I could use more help on the micropipettes and what the amounts on the front with the numbers pertain to. I think we could use a little more practice on making solutions and adjusting pH may be on more time would assure our skills.
We eventually, redesigned and successfully did our experiment to see results. The DNA strands were really cool to see with the gel process. I was happy we didn't really do all this work for nothing. Even though we didn't do a whole lot of work it was nice to see some results after a chemical error.