Purpose: To determine the frequency of the Alu insert in the STEM Bio class using PCR.
Hypothesis: If there are no results, then I will ask the teacher for help to redo the experiment for some results. This is not a real lab without some results, but I guess no results can be a result as well.
Materials:
Cheek cells
Chelex
Master Mix : nucleotides, DNA Polymorase
Primer Mix
TBE Buffer, Agarose gel
DNA stain solution
Procedure: To get to our final result, we had to go through many steps. First, we swirled and swished a saline solution around in our mouths and spit it out in the cup after 30 seconds. We used the micropipets to take some of the DNA and put it in a test tube with some reagents. We put the tubes in a centrifuge and let it spin for 1 or 2 seconds, When we took them out, it appeared that there were little cell pellets at the bottom of each tube (if you did it right). We poured out most of the liquid above the pellet, and then withdrew some of the remaining liquid into a different tube with Chelex in it. We heated the tubes up for about 10 minutes. After releasing the pressure from the small tube, we put it in the centrifuge for 1 minute. Then, we took a smaller tube and put some of our Chelex/DNA in it and refrigerated it for about 24 hours.
We obtained a tiny PCR tube and put some Master Mix, Primer Mix, and DNA in it. We put it in the microcentrifuge for 10 seconds, and added loading dye to it afterwards. Then, we carefully pipetted the mixture into a slit in the agarose gel; the dye was green. Then some bp ladder was added to each well and electrophoresed the samples for 20-45 minutes. After the electrophoresis, the gels could be stained and photographed.
Hypothesis: If there are no results, then I will ask the teacher for help to redo the experiment for some results. This is not a real lab without some results, but I guess no results can be a result as well.
Materials:
Cheek cells
Chelex
Master Mix : nucleotides, DNA Polymorase
Primer Mix
TBE Buffer, Agarose gel
DNA stain solution
Procedure: To get to our final result, we had to go through many steps. First, we swirled and swished a saline solution around in our mouths and spit it out in the cup after 30 seconds. We used the micropipets to take some of the DNA and put it in a test tube with some reagents. We put the tubes in a centrifuge and let it spin for 1 or 2 seconds, When we took them out, it appeared that there were little cell pellets at the bottom of each tube (if you did it right). We poured out most of the liquid above the pellet, and then withdrew some of the remaining liquid into a different tube with Chelex in it. We heated the tubes up for about 10 minutes. After releasing the pressure from the small tube, we put it in the centrifuge for 1 minute. Then, we took a smaller tube and put some of our Chelex/DNA in it and refrigerated it for about 24 hours.
We obtained a tiny PCR tube and put some Master Mix, Primer Mix, and DNA in it. We put it in the microcentrifuge for 10 seconds, and added loading dye to it afterwards. Then, we carefully pipetted the mixture into a slit in the agarose gel; the dye was green. Then some bp ladder was added to each well and electrophoresed the samples for 20-45 minutes. After the electrophoresis, the gels could be stained and photographed.