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ETD 18: Draw + Explain Both: A. How you would make tons of EzFLOW Protein? B. How does Gel Electrophoresis work to solve crimes?
October 17, 2013 at 3:14pm

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Amber Neathery- Period 3
October 28, 2015 at 9:48pm

Parent: a) I learned that the making of easy flow protein which is an anti clotting agent starts from the saliva of leeches, where a protein can be found.   the process of isolating the anticoagulant protein includes:  plasmid, restriction enzyme, ligase, and recombinant dna,  bacteria growing, popping membrane with alcohol, and centrifuging.  

b) by starting with  restricting enzyme that cuts dna in certain spots.  now dna is in fragment which is placed in a well with a positive and negative end.   since bigger sections of dna gravitate to the top in comparison to the smaller segments,  identification of suspected individual is possible.   

Student: Teaching my dad about making the EZ Flow protein allowed me to makes sense of the first four steps of the ETD! I realized that the plasmid needed to be cut and the phosphate backbone rebound by ligase in order to place the gene of interest just downstream of the lac Z. I understood the process of using gel electrophoresis to solve crime because of the related homework assignment, but explaining it again emphasized the use of restriction enzymes for me. 

Anne Sweeney
December 09, 2015 at 7:00pm

Parent:Anne did a good job explaining how more proteins can be created using plasmids. It’s quite an intricate process.  I thought it was interesting how they use gel electrophoresis  to compare DNA samples when solving criminal cases.

Student: When explaining the process of how plasmids are used to make ezflow protein, I realized I needed to define common vocabulary for my mom. It took longer than I expected to explain because I needed to cover what some of the words meant in order for her to understand the process as a whole. Gel Electrophoresis was easier to explain when I drew out a diagram and made it a step by step process. This helped me review the basic concepts I needed to know from this ETD.

Anne Sweeney
December 27, 2015 at 3:04pm

Parent: Anne did a good job explaining how more proteins can be created using plasmids. It’s quite an intricate process. I thought it was interesting how they use gel electrophoresis  to compare DNA samples when solving criminal cases.

Student: When explaining the process of how plasmids are used to make ezflow protein, I realized I needed to define common vocabulary for my mom. It took longer than I expected to explain because I needed to cover what some of the words meant in order for her to understand the process as a whole. Gel Electrophoresis was easier to explain when I drew out a diagram and made it a step by step process. This helped me review the basic concepts I needed to know from this ETD.

Kate Freeman
January 19, 2016 at 10:41pm

Parent: I learned that gel electrophoresis is a method for separation and analysis of macromolecules and their fragments based on size and shape. I found the process very interesting and useful in the method of solving crimes. Kate also taught me that there is an easy and efficient way to produce tons of EZ flow protein by taking genes and inserting them into an existing plasmid.

Student: I explained to my dad that in order to produce large amounts of EZ flow protein you must take an existing plasmid then use the same restriction enzymes on all of the new proteins to cut out all of the sticky ends and finally, use ligase to link them all together and form a recombinant plasmid. I then discussed how police use gel electrophoresis by collecting DNA and amplifying it using PCR, then using the same restriction enzyme to cut the ECORI, and finally running gel DNA which shows negative change and seperates pusler DNA by size. Reviewing these processes helped me remember that restriction enzymes cut genes and can be used for cloning.

Alyssa Hajek
October 27, 2016 at 10:14pm

Parent: She explained to me the 8 steps of how EZ flow protein is made. Very complex process for sure.I thought the leach concept was really helpful start to the lesson.  She also drew a diagram to demonstrate gel electrophoresis.  Very nice lesson on a complicated subject and I suspect i will not retain much of it.

 

Student: I explained to my dad the steps you would take if you were trying to mass produce the EZ flow protein.  While explaining this to him I had to explain more in depth what different things mean.  For example I had to explain to him what a restriction enzyme does and why it is so important to use the same restriction enzyme so you make the same cuts.  I now have a better understanding of the process of gene expression.  I also told my dad about gel electrophoresis.  I told him, again, about the importance of using the same restriction enzyme. I made it clear to him that the length of the piece of DNA decides how far it will travel in the gel.

Jacob Michaels
January 03, 2018 at 7:00pm

Parent-  EZ flow protein is an anti-clotting protein found in leeches. If you wanted to make a large quantity of it you would need to identify the gene of interest and use restriction enzymes to cut out a portion of the DNA and put a plasmid in its place, closing it up with ligase.  To do that you would need to use CACL2, heat shock and ice to manipulate the cell membrane and slip the plasmid inside.  Once it is in, reproduction of cells can be sped up to make large quantities of the protein.

Gel electrophoresis can solve crimes by looking at fragment lengths of DNA.  One can use restriction enzymes to cut the DNA at restriction sites, then run it through the gel from negative to positive.  It pushes the DNA, separating fragments by size for comparison.

Student- Making EZ flow protein is a complex process but I believe I summed it up OK for my dad, he seemed to understand it pretty well. The vocabulary tripped me up a bit but I looked back through the ETD and remembered what everything was. Gel Electrophoresis was easier to explain and understand like using restriction enzymes at restriction sites and how far each fragment goes is based on size.

Sarah Kropelnicki
January 27, 2019 at 6:24pm

Parent: Today I learned that EZ flow is hirudin which is an anti-clotting agent in leeches. To produce mass amounts of it, you would first need to find the existing plasmid with said anti-clotting agent within it. Then, through the use of restriction enzymes, which cut DNA like a pair of scissors. After this, the DNA of the ampR and the EZ flow is left with sticky ends which are glued together with ligase. You then have a recombinant plasmid, through which bacterial transformation selects the ampR. After incubating, you add lactose to make the EZ flow protein. Finally, you pop the cells to get the EZ flow out of the bacteria. 

Student: As I taught this, I was able to connect more dots as we have just learned more about ligase in DNA replication. Also as I went through gel electrophoresis, I explained that one must collect DNA from the persons of interest, use the same restriction enzyme to cut a certain place in each DNA, and then run the gel. The DNA, which is negative, will run negative to positive, the smaller fragments traveling farther than the longer fragments. This creates a kind of picture of DNA, which can be compared to another and identified. 

Alaina Brady
October 22, 2019 at 11:33pm

Parent: I learned one way in which drug companies make money, isolating and selling an easy flow protein to prevent and treat blood clots.  It starts with a leech which contains a protein called hinadin, that thins it's victims blood so that the leech can suck more blood.  You use leech DNA and cut it around the around the code for the hinadin and then use ligase to bind the gene to the plasmid.  You also use an ori, which helps the plasmid replicate. You also add  a resistance gene that will help the later used bacteria survive if the plasmid gets reproduced. An operon is also needed to start the process.  This is now called a recombinate plasmid.  You then made a bunch of the stuff and put it in bacteria which reproduces creating a large source of the plasmid within bacteria that carry it.  Then you pop the bacteria cell and centrifuge that substance to be able extract the easy flow protein.

Alaina also told me about a machine called gel electrophoresis that is used in helping identify matching strands of DNA identify suspects.  The machine sends an electric current thru the gel which contains the different DNA samples and this sort the DNA samples by the sizes in which they are broken, with the smallest on the bottom.  Prior to placing the DNA in the gel, the restriction enzyme or "cutter" is used to cut each DNA sample and as each sample is unique, it is cut in a unique manner.  So you'd get DNA and from the crime scene and from your suspect and run each through the process,  The guilty suspect will make exactly to the control sample as it is cut the same way every time, and everyone'2s is cut differently

Student: I told my mom how to make a recombinant plasmid to produce a bunch of EZ flow protein. I taught her that you put in your gene of interest, an ori, a resistance gene, and an operon in the plasmid. You then insert that plasmid into bacteria with a substance that will kill the bacteria unless it carries the plasmid with the resistance gene. This selects for the bacteria that carries the plasmid. Then that bacteria can reproduce and then you have access to a bunch of EZ flow. So you pop the cell and centrifuge it to sort the different substances by density. Then you have your EZ flow protein.

I also told my mom how to solve crime. You collect your crime scene DNA and your suspect DNA. Then you cut each set of DNA with the same restriction enzyme, and different lengths of DNA strands are formed. You then run those strands through gel electrophoresis, which sorts the strands by length, longest to shortest. The suspect with the DNA that perfectly matches the Crime Scene DNA committed the crime.

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