Purpose:
the purpose of this lab was to make 10 milliliters (mL) of 5 NaCl solution, and also to make 100mL of TE buffer from 10 mM TRIS, 1 mM EDTA. then we had to find out if DNA could be spooled out of solution, to find out what DNA looks like and its many unique properties, and Finally we needed to find out what yield of DNA can be recovered during the isolation, to prepare and pour an agarose gel for DNA fragment analysis, to find out what the appearance of different DNA samples on an agarose gel.
Materials List:
Lab 4A
Lab 4B
Lab 4I
Lab 4J
Procedure 4a
Data analysis:
Our experiment failed and we did not get any results, sadly. our bands did not show up. After talking about it to Dr. LB we came to the conclusion that our stain might have been susceptible to UV rays over the year of being left out. The only way to actually find out is if we hand made the stain and use the new stain to test the tissue. another reason could have been human error but that was highly unlikely because it would have only ruined some of the experiments, bot all of them.
conclusion:
Isolating DNA can also be called purifying the DNA and it is very important, because it helps forensic scientists find the finger print of the coulpret. the use of DNA can help scientists study a specific gene. The gene can help scientists determine a treatment for a particular desease. Scientists use DNA to help studies, like insulin, hormones, and antibodies.
reflection:
My group was very diligent at working on this project and even though we didn't get the results we wanted, we had fun trying and would have succeeded if the stain wasn't left out in the lab for so long. my group was Rosalie, Cole, Zach, Tony, Gabby, and Ingrid. We worked together pretty well and got stuff done. My group was very successful in making all the solutions and adjusting the pH level to what it needed to be. There was only one person in our group that couldn't pipet very well but instead of doing it for them we just taught them how to do it better, which pushed us further as a group.
the purpose of this lab was to make 10 milliliters (mL) of 5 NaCl solution, and also to make 100mL of TE buffer from 10 mM TRIS, 1 mM EDTA. then we had to find out if DNA could be spooled out of solution, to find out what DNA looks like and its many unique properties, and Finally we needed to find out what yield of DNA can be recovered during the isolation, to prepare and pour an agarose gel for DNA fragment analysis, to find out what the appearance of different DNA samples on an agarose gel.
Materials List:
Lab 4A
- analytical balance
- tabletop milligram balance
- weigh paper
- weigh boat
- lab scoops
- sodium chloride
- 15 ml tubes
- tube racks for 15 ml tubes
- TRIS
- EDTA
- 125 ml bottle
- 100 ml graduated cylinder
- pH paper, wide/narrow-range
- Hydrochloric acid
- Sodium hydroxide
- glass rods
Lab 4B
- 50 ml beakers
- salmon sperm DNA
- 2 - 20 ml pipet
- pipet pump
- micropipet (P-1000)
- micropipet tips for P-1000
- ethanol (95%)
- glass rods
- 15 ml conical tubes (capped)
- tube racks fro 15 ml conical tubes
- permanent lab markers
Lab 4I
- TAE buffer concentrate (40x stock)
- 600 ml beaker
- agarose
- tabletop milligram balance
- weigh paper
- lab scoops
- 250 ml media bottle
- permanent lab markers
- microwave oven
- hot hands protector
- horizontal gel box for agarose gels
- 50 ml beakers
Lab 4J
- horizontal Gel box for agarose gels
- prepared agarose gel
- TAE buffer concentrate (40x stock)
- tube rack for 15 ml conical tubes
- permanent lab markers
- DNA samples
- gel loading dye (6x)
- micropipet (P-2-20)
- micropipet (P-20-200)
- micropipet tips for P-2-20
- micropipet tips for P-20-200
- microcentrifuge
- power supply
- ethidium bromide (0.5 micrograms/ml)
- gloves
Procedure 4a
- 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
Data analysis:
Our experiment failed and we did not get any results, sadly. our bands did not show up. After talking about it to Dr. LB we came to the conclusion that our stain might have been susceptible to UV rays over the year of being left out. The only way to actually find out is if we hand made the stain and use the new stain to test the tissue. another reason could have been human error but that was highly unlikely because it would have only ruined some of the experiments, bot all of them.
conclusion:
Isolating DNA can also be called purifying the DNA and it is very important, because it helps forensic scientists find the finger print of the coulpret. the use of DNA can help scientists study a specific gene. The gene can help scientists determine a treatment for a particular desease. Scientists use DNA to help studies, like insulin, hormones, and antibodies.
reflection:
My group was very diligent at working on this project and even though we didn't get the results we wanted, we had fun trying and would have succeeded if the stain wasn't left out in the lab for so long. my group was Rosalie, Cole, Zach, Tony, Gabby, and Ingrid. We worked together pretty well and got stuff done. My group was very successful in making all the solutions and adjusting the pH level to what it needed to be. There was only one person in our group that couldn't pipet very well but instead of doing it for them we just taught them how to do it better, which pushed us further as a group.