Introduction
a. Background- This lab simulates what happens when bodily fluids are shared and how ELISA is used to determine if you have been exposed to a disease. ELISA uses antibodeis to detect the precense of an intigen in the body and track it back to its source. In ELISA, antibodies bind to wells in a gel and antibodies are added and attatch to their corresponding antigens and then when a substrate is added, the wells will change colors and determine if the individual has a disease.
b. Purpose- The purpose of this lab is to show how ELISA works and how it could be applied to real life situations. ELISA is the scientific process that is used in tests that determine if someone has a disease or if a woman is pregnant. It has also been used in testing for drug use and testing air quality. The test is useful because it gives a distinct positive or negative result.
c. Procedure- On Day 1, we will transfer bodily fluids that we shared with another individual into test tubes and mix the samples. On Day 2, we will pipet the samples into a 12-well strip with weals designated for positive and negative controls. We will transfer 50 microliters into each well and then wait until the proteins in the sample bind to the bottoms of the wells. After we will tip the strip upside down and fill each well with wash buffer. Then we will transfer primary antibody into the wells and wait for the antibodies to bind to their targets. We will again wash the strip twice. Then we will add secondary antibodies into the wells and then wait for them to bind to their targets. Then yo will wash the wells again three times. AFte we will transfer enzyme substrate into the wells and after waiting five minutes, we will record the results as some of the wells will change colros. Finally, we will clean up.
d. Hypothesis- I hypothosize that at least one of the members at our lab table will have the diseas4e.
Monday, May 16, 2011
Thursday, April 21, 2011
I Got It From My Mama: Mitochondrial DNA Lab
Introduction
a. Background- In this lab, we will be testing mitochondrial DNA to see if an individual carries a gene for a disease. We receive our mitochondrial DNA from our moms. In order to do so, one must test the genome, which is our hereditary code. We will be extracting our own mitochondrial DNA and mass producing in through PCR (Polymerase Chain Reaction). The necessary components for PCR are primers, nucleotides, DNA polymerase, and the existing DNA template to be replicated. PCR is beneficial because it can replicate large amounts of DNA that were formerly not able to be replicated. After replicating the gene, we will run the DNA through a gel, which will show us if we carry the gene for the disease.
b. Purpose- This procedure is important because it could be used to detect if one carries the genes for a disease before symptoms actually occur. This could mean lives and large amounts of money being saved by early detection of diseases. It is also important because it could help us determine if we got a disease-causing gene from our mothers.
c. Summary- On Day 1 of the lab, we will be extracting our mitochondrial DNA. first we will collect DNA from our cheek cells and break open the cell membranes. We will also add instagene matrix to kill DNAse, which could harm the DNA. On Day 2, we will be using PCR to replicate the small amount of DNA that we extracted. Primers used in PCR target the "disease gene" in the genome. On Day 3, we use gel electrophoresis to diagram the results of the test.
d. Hypothesis- I hypothesize that one of us will have the "disease gene".
Procedure
On day 1, we rinsed our mouths vigorously with saline for 30 seconds. We transfered 1 ml of the saline rinse into a micro test tube containing 200 microliters of InstaGene matrix. We then centrifuged the tubes for 2 minutes, and saw a pellet of cell appear at the bottom of the tube. After pelleting the cells, we poured off the saline and re suspended the pellet by vortexing the tube. Next we micropipetted 20 microliters of of cells into the tube containing InstaGene, and then again vortexed the tube. Then we incubated the tube at 56 degrees Celsius for ten minutes, but at 5 minutes we vortexed the tubes before returning them back to the bath. After, we placed the tubes in a boiling water bath for 5 minutes. Then, we vortexed the contents for 5 minutes and refrigerated the tubes overnight. On day 2, we centrifuged the tubes for 2 minutes. Then we transferred 20 microliters of the DNA template into the PCR tube being sure to not transfer any matrix beads. We also added 20 microliters of the master mix into the PCR tube. Finally we placed the tubes into the thermal cycler. On day 3, we obtained our PCR tubes and centrifuged them for 3 seconds. Then we added 10 microliters of loading dye into the PCR tube and mixed it gently. After we pipeted 10 microliters of the marker into lane 1 of the gel. In lanes 2, 3, 4, and 5, we each placed 20 microliters of our mitochondrial DNA. After we ran the gel at 200 volts for 3 minutes. When the gel was complete, it was stained and then we analyzed it the next day.
Results
Discussion
After analyzing our results, it turns out that I do not have the "disease" that we were testing for. Although there were no obvious errors in our lab, one source of error could be pipetting DNA as well as matrix beads into the PCR tube and then having the DNA be destroyed.
a. Background- In this lab, we will be testing mitochondrial DNA to see if an individual carries a gene for a disease. We receive our mitochondrial DNA from our moms. In order to do so, one must test the genome, which is our hereditary code. We will be extracting our own mitochondrial DNA and mass producing in through PCR (Polymerase Chain Reaction). The necessary components for PCR are primers, nucleotides, DNA polymerase, and the existing DNA template to be replicated. PCR is beneficial because it can replicate large amounts of DNA that were formerly not able to be replicated. After replicating the gene, we will run the DNA through a gel, which will show us if we carry the gene for the disease.
b. Purpose- This procedure is important because it could be used to detect if one carries the genes for a disease before symptoms actually occur. This could mean lives and large amounts of money being saved by early detection of diseases. It is also important because it could help us determine if we got a disease-causing gene from our mothers.
c. Summary- On Day 1 of the lab, we will be extracting our mitochondrial DNA. first we will collect DNA from our cheek cells and break open the cell membranes. We will also add instagene matrix to kill DNAse, which could harm the DNA. On Day 2, we will be using PCR to replicate the small amount of DNA that we extracted. Primers used in PCR target the "disease gene" in the genome. On Day 3, we use gel electrophoresis to diagram the results of the test.
d. Hypothesis- I hypothesize that one of us will have the "disease gene".
Procedure
On day 1, we rinsed our mouths vigorously with saline for 30 seconds. We transfered 1 ml of the saline rinse into a micro test tube containing 200 microliters of InstaGene matrix. We then centrifuged the tubes for 2 minutes, and saw a pellet of cell appear at the bottom of the tube. After pelleting the cells, we poured off the saline and re suspended the pellet by vortexing the tube. Next we micropipetted 20 microliters of of cells into the tube containing InstaGene, and then again vortexed the tube. Then we incubated the tube at 56 degrees Celsius for ten minutes, but at 5 minutes we vortexed the tubes before returning them back to the bath. After, we placed the tubes in a boiling water bath for 5 minutes. Then, we vortexed the contents for 5 minutes and refrigerated the tubes overnight. On day 2, we centrifuged the tubes for 2 minutes. Then we transferred 20 microliters of the DNA template into the PCR tube being sure to not transfer any matrix beads. We also added 20 microliters of the master mix into the PCR tube. Finally we placed the tubes into the thermal cycler. On day 3, we obtained our PCR tubes and centrifuged them for 3 seconds. Then we added 10 microliters of loading dye into the PCR tube and mixed it gently. After we pipeted 10 microliters of the marker into lane 1 of the gel. In lanes 2, 3, 4, and 5, we each placed 20 microliters of our mitochondrial DNA. After we ran the gel at 200 volts for 3 minutes. When the gel was complete, it was stained and then we analyzed it the next day.
Results
Discussion
After analyzing our results, it turns out that I do not have the "disease" that we were testing for. Although there were no obvious errors in our lab, one source of error could be pipetting DNA as well as matrix beads into the PCR tube and then having the DNA be destroyed.
Monday, April 11, 2011
"Fish are friends, not food”: Using Proteomics to Determine Relatedness Between Fish Species
Introduction
a. Background: In this lab, we will be using proteomics to determine the relatedness between species. While a single gene can encode multiple proteins, to what level and where a protein is express or changed such protein after it is translated can result in proteins with very differnent functions. The study of protein structure and functions is called proteomics. The proteome differs from cell to cell and is constantly changing. It is hypothesized that the noncoding DNA controls and regulates protein production. Currently, the study of proteins and their functions are being coordinated by the Human Proteome Organization.
b. Purpose: The purpose of this lab is to compare the proteins of multiple species of fish to determine their relatedness. Proteomics is used to compare other species based on factors beyond their genomes.
c. Process: First we will take multiple species of fish and denature their muscle. By doing so, we will be unraveling the structures of the actin and myosin proteins. Next we will pipet the samples into a protein gel and do gel electrophoresis. After, we will observe the gel results to determine the relatedness between species. The more similar band patterns species have, the more related they are.
d. Hypothesis: I hypothesize that we will find species of fish that are related to one another.
Procedure
Results
a. Background: In this lab, we will be using proteomics to determine the relatedness between species. While a single gene can encode multiple proteins, to what level and where a protein is express or changed such protein after it is translated can result in proteins with very differnent functions. The study of protein structure and functions is called proteomics. The proteome differs from cell to cell and is constantly changing. It is hypothesized that the noncoding DNA controls and regulates protein production. Currently, the study of proteins and their functions are being coordinated by the Human Proteome Organization.
b. Purpose: The purpose of this lab is to compare the proteins of multiple species of fish to determine their relatedness. Proteomics is used to compare other species based on factors beyond their genomes.
c. Process: First we will take multiple species of fish and denature their muscle. By doing so, we will be unraveling the structures of the actin and myosin proteins. Next we will pipet the samples into a protein gel and do gel electrophoresis. After, we will observe the gel results to determine the relatedness between species. The more similar band patterns species have, the more related they are.
d. Hypothesis: I hypothesize that we will find species of fish that are related to one another.
Procedure
Results
Tuesday, March 15, 2011
Catch My Disease: Disease Gene Lab
Introduction
a. Background- In this lab, we will be testing DNA to see if an individual carries a gene for a disease. In order to do so, one must test the genome, which is our hereditary code. We will be extracting our own DNA and mass producing in through PCR (Polymerase Chain Reaction). The necessary components for PCR are primers, nucleotides, DNA polymerase, and the existing DNA template to be replicated. DNA is beneficial because it can replicate large amounts of DNA that were formerly not able to be replicated. After replicating the gene, we will run the DNA through a gel, which will show us if we carry the gene for the disease.
b. Purpose- This procedure is important because it could be used to detect if one carries the genes for a disease before symptoms actually occur. This could mean lives and large amounts of money being saved by early detection of diseases.
c. Summary- On Day 1 of the lab, we will be extracting DNA. first we will collect DNA from our cheek cells and break open the cell membranes. We will also add instagene matrix to kill DNAse, which could harm the DNA. On Day 2, we will be using PCR to replicate the small amount of DNA that we extracted. Primers used in PCR target the "disease gene" in the genome. On Day 3, we use gel electrophoresis to diagram the results of the test.
d. Hypothesis- I hypothesize that one of us will have the "disease gene".
Procedure
On day 1, we rinsed our mouths vigorously with saline for 30 seconds. We transfered 1 ml of the saline rinse into a micro test tube containing 200 microliters of InstaGene matrix. We then centrifuged the tubes for 2 minutes, and saw a pellet of cell appear at the bottom of the tube. After pelleting the cells, we poured off the saline and re suspended the pellet by vortexing the tube. Next we micropipetted 20 microliters of of cells into the tube containing InstaGene, and then again vortexed the tube. Then we incubated the tube at 56 degrees Celsius for ten minutes, but at 5 minutes we vortexed the tubes before returning them back to the bath. After, we placed the tubes in a boiling water bath for 5 minutes. Then, we vortexed the contents for 5 minutes and refrigerated the tubes overnight. On day 2, we centrifuged the tubes for 2 minutes. Then we transferred 20 microliters of the DNA template into the PCR tube being sure to not transfer any matrix beads. We also added 20 microliters of the master mix into the PCR tube. Finally we placed the tubes into the thermal cycler. On day 3, we obtained our PCR tubes and centrifuged them for 3 seconds. Then we added 10 microliters of loading dye into the PCR tube and mixed it gently. After we pipeted 10 microliters of the marker into lane 1 of the gel, homozygous (+/+) control in lane 2, homozygous (-/-) control in lane 3, and heterozygous (+/-) in lane 4. In lanes 5, 6, 7, and 8, we each placed 20 microliters of our DNA. After we ran the gel at 200 volts for 3 minutes. When the gel was complete, it was stained and then we analyzed it the next day.
Results
Discussion
After analyzing our results, it turns out that I do not have the "disease" that we were testing for. Although there were no obvious errors in our lab, one source of error could be pipetting DNA as well as matrix beads into the PCR tube and then having the DNA be destroyed.
a. Background- In this lab, we will be testing DNA to see if an individual carries a gene for a disease. In order to do so, one must test the genome, which is our hereditary code. We will be extracting our own DNA and mass producing in through PCR (Polymerase Chain Reaction). The necessary components for PCR are primers, nucleotides, DNA polymerase, and the existing DNA template to be replicated. DNA is beneficial because it can replicate large amounts of DNA that were formerly not able to be replicated. After replicating the gene, we will run the DNA through a gel, which will show us if we carry the gene for the disease.
b. Purpose- This procedure is important because it could be used to detect if one carries the genes for a disease before symptoms actually occur. This could mean lives and large amounts of money being saved by early detection of diseases.
c. Summary- On Day 1 of the lab, we will be extracting DNA. first we will collect DNA from our cheek cells and break open the cell membranes. We will also add instagene matrix to kill DNAse, which could harm the DNA. On Day 2, we will be using PCR to replicate the small amount of DNA that we extracted. Primers used in PCR target the "disease gene" in the genome. On Day 3, we use gel electrophoresis to diagram the results of the test.
d. Hypothesis- I hypothesize that one of us will have the "disease gene".
Procedure
On day 1, we rinsed our mouths vigorously with saline for 30 seconds. We transfered 1 ml of the saline rinse into a micro test tube containing 200 microliters of InstaGene matrix. We then centrifuged the tubes for 2 minutes, and saw a pellet of cell appear at the bottom of the tube. After pelleting the cells, we poured off the saline and re suspended the pellet by vortexing the tube. Next we micropipetted 20 microliters of of cells into the tube containing InstaGene, and then again vortexed the tube. Then we incubated the tube at 56 degrees Celsius for ten minutes, but at 5 minutes we vortexed the tubes before returning them back to the bath. After, we placed the tubes in a boiling water bath for 5 minutes. Then, we vortexed the contents for 5 minutes and refrigerated the tubes overnight. On day 2, we centrifuged the tubes for 2 minutes. Then we transferred 20 microliters of the DNA template into the PCR tube being sure to not transfer any matrix beads. We also added 20 microliters of the master mix into the PCR tube. Finally we placed the tubes into the thermal cycler. On day 3, we obtained our PCR tubes and centrifuged them for 3 seconds. Then we added 10 microliters of loading dye into the PCR tube and mixed it gently. After we pipeted 10 microliters of the marker into lane 1 of the gel, homozygous (+/+) control in lane 2, homozygous (-/-) control in lane 3, and heterozygous (+/-) in lane 4. In lanes 5, 6, 7, and 8, we each placed 20 microliters of our DNA. After we ran the gel at 200 volts for 3 minutes. When the gel was complete, it was stained and then we analyzed it the next day.
Results
Discussion
After analyzing our results, it turns out that I do not have the "disease" that we were testing for. Although there were no obvious errors in our lab, one source of error could be pipetting DNA as well as matrix beads into the PCR tube and then having the DNA be destroyed.
Tuesday, February 1, 2011
Is My Strawberry a Frankenfood?: GMO Foods Lab
Introduction
a) Background- GMO's are genetically modified organisms, and can be used in various scientific settings, often in agriculture, such as in the development of making genetically modified foods, that could be resistant to disease or frost or having a higher amount of vitamins in it. GMO's are made by taking a plasmid with the gene of interest (a tumor inducing plasmid) and putting it into agro bacterium. Then we will place the agro bacterium in a plant cell. The plasmic will take the DNA and will transcribe it, creating a genetically modified plant. GMO's are identified throught two techniques, ELISA and PCR. We are using PCR in this lab, where we identify sequences of DNA that have been inserted into the GM plant. DNA fragments can be isolated from highly processed foods. GMO's are controversial because some people identify genetically modified organisms as "unnatural" and have the potential to create organisms that will be resistant to normal toxins. Many also worry about potential allergic reactions to GMO's and think that organic foods are better for the environment.
b) Purpose- The purpose of this lab is to take an organism and determine if it is genetically modified or not by comparing it to a known organic organism. The technique of PCR is used regularly in the scientific world today, for example in forensics and genetic studies.
c) Procedure- First we will mortar and pestal the cell of an organism to break down its cell wall. Then we will place the crushed cell in a 99 degree Celcius water bath to break down the nuclear and cell membranes. we will also have to add Instagene Matrix beads that kill DNAse enzymes, otherwise the DNAse will come into contact with the DNA because the cell wall is broken and will destroy it. Next, we will use the PCR process using two primers, one to target plant DNA (the control) and one to target GM DNA. Finally, we will use gel electrophoresis to visualize the gene of interest and determine the presence/absence of the desired PCR products.
d) Hypothesis- In this lab we are testing if the organism we bring in is genetically modified or not.
Procedure
On Day 1, we will label two tubes with Test Food 1 and Test Food 2. After we weighed out about 0.9 grams of non-GMO food (corn flour) and put it in the mortar, along with 10 milliliters of distilled water. We ground up this mixture for around two minutes and then followed by adding 5 milliliters of distilled water. Next we pipetted 50 microliters of the slurry into the tube containing InstaGene. After, we prepared our tube for our second test food, strawberries, using the same procedure. Then we flicked both tubes and placed the tubes in a 95 degree Celsius water bath for 5 minutes. We followed by placing the tubes in a centrifuge for 5 minutes and then we refrigerated the tubes over night. On Day 2, we labeled six PCR tubes. Tubes 1 and 2 contained 20 microliters of non-GMO food control DNA. Tubes 3 and 4 contained 20 microliters of test food DNA. Tubes 5 and 6 contained 20 microliters of GMO positive control DNA. After we labeled these tubes, we placed them in the foam float on ice. Then we added 20 microliters of plant master mix to tubes 1, 3, and 5 and 20 microliters of GMO master mix to tubes 2, 4, and 6. Finally we placed the PCR tubes in the thermal cycler. On Day 3, we obtained our PCR tube from the thermal cycler and centrifuged it for three seconds. Next we pipetted 10 microliters of Orange G loading dye to test tubes 1-6. Then we loaded 20 microliters of each sample (in the tubes) into rows 1-6 of our gel. In row 7, Mr. Chugh loaded 20 microliters of PCR molecular weight ruler, to act as a control for our lab. Then we ran the gel for 3 minutes at 200 volts. The se stained in Fast Blast DNA stain. The next day, we observed our results.
Results
We concluded that our strawberries were genetically modified.
Discussion
When observing our gel the next day, we found banding pattern in rows 3, 4, 5 and 6. Because there was a band in lane 4 of the gel, we can conclude that the strawberries we tested were genetically modified. One possible source of error for this lab was not mixing the loading dye well in each sample. Other sources of error could be that PCR or extraction could have been unsuccessful.
a) Background- GMO's are genetically modified organisms, and can be used in various scientific settings, often in agriculture, such as in the development of making genetically modified foods, that could be resistant to disease or frost or having a higher amount of vitamins in it. GMO's are made by taking a plasmid with the gene of interest (a tumor inducing plasmid) and putting it into agro bacterium. Then we will place the agro bacterium in a plant cell. The plasmic will take the DNA and will transcribe it, creating a genetically modified plant. GMO's are identified throught two techniques, ELISA and PCR. We are using PCR in this lab, where we identify sequences of DNA that have been inserted into the GM plant. DNA fragments can be isolated from highly processed foods. GMO's are controversial because some people identify genetically modified organisms as "unnatural" and have the potential to create organisms that will be resistant to normal toxins. Many also worry about potential allergic reactions to GMO's and think that organic foods are better for the environment.
b) Purpose- The purpose of this lab is to take an organism and determine if it is genetically modified or not by comparing it to a known organic organism. The technique of PCR is used regularly in the scientific world today, for example in forensics and genetic studies.
c) Procedure- First we will mortar and pestal the cell of an organism to break down its cell wall. Then we will place the crushed cell in a 99 degree Celcius water bath to break down the nuclear and cell membranes. we will also have to add Instagene Matrix beads that kill DNAse enzymes, otherwise the DNAse will come into contact with the DNA because the cell wall is broken and will destroy it. Next, we will use the PCR process using two primers, one to target plant DNA (the control) and one to target GM DNA. Finally, we will use gel electrophoresis to visualize the gene of interest and determine the presence/absence of the desired PCR products.
d) Hypothesis- In this lab we are testing if the organism we bring in is genetically modified or not.
Procedure
On Day 1, we will label two tubes with Test Food 1 and Test Food 2. After we weighed out about 0.9 grams of non-GMO food (corn flour) and put it in the mortar, along with 10 milliliters of distilled water. We ground up this mixture for around two minutes and then followed by adding 5 milliliters of distilled water. Next we pipetted 50 microliters of the slurry into the tube containing InstaGene. After, we prepared our tube for our second test food, strawberries, using the same procedure. Then we flicked both tubes and placed the tubes in a 95 degree Celsius water bath for 5 minutes. We followed by placing the tubes in a centrifuge for 5 minutes and then we refrigerated the tubes over night. On Day 2, we labeled six PCR tubes. Tubes 1 and 2 contained 20 microliters of non-GMO food control DNA. Tubes 3 and 4 contained 20 microliters of test food DNA. Tubes 5 and 6 contained 20 microliters of GMO positive control DNA. After we labeled these tubes, we placed them in the foam float on ice. Then we added 20 microliters of plant master mix to tubes 1, 3, and 5 and 20 microliters of GMO master mix to tubes 2, 4, and 6. Finally we placed the PCR tubes in the thermal cycler. On Day 3, we obtained our PCR tube from the thermal cycler and centrifuged it for three seconds. Next we pipetted 10 microliters of Orange G loading dye to test tubes 1-6. Then we loaded 20 microliters of each sample (in the tubes) into rows 1-6 of our gel. In row 7, Mr. Chugh loaded 20 microliters of PCR molecular weight ruler, to act as a control for our lab. Then we ran the gel for 3 minutes at 200 volts. The se stained in Fast Blast DNA stain. The next day, we observed our results.
Results
We concluded that our strawberries were genetically modified.
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Discussion
When observing our gel the next day, we found banding pattern in rows 3, 4, 5 and 6. Because there was a band in lane 4 of the gel, we can conclude that the strawberries we tested were genetically modified. One possible source of error for this lab was not mixing the loading dye well in each sample. Other sources of error could be that PCR or extraction could have been unsuccessful.
Tuesday, January 25, 2011
GTL: Gene Transformation Lab
Introduction
a) Background- In this lab, we will be using a process known as genetic transformation, where we cause a change in an organism by inserting a gene into an organism. Genetic transformation has been used in many sectors of biotechnology. In farming, it has been used to foster resistance to frost, spoilage, and pests. In bioremediation, bacteria can be transformed to digest oil, which is useful in the cleanup of oil spills. In medicine, diseases caused by malfunctioning genes are now being able to be transformed in gene therapy. This could potentially mean that a person's cells that are making them sick could be transformed with healthy copies.
b) Purpose- The purpose of this lab is to show how by inserting a gene into an organism, we can cause a change in that organism. In this instance, we would be transforming a bacteria with a gene that codes for Green Fluorescent Protein that will cause the bacteria to fluoresce under ultraviolet light.
c) Process- First we are labeling one closed test tube +pGLO and another-pGLO. Then we pipet 250 microliters of transformation solution into the tubs and place them on ice. Using a loop, we pick up the +pGLO immerse the solution at the bottom of the tube and then do the same with -pGLO. Then we add 10 microliters of plasmid DNA directly to the liquid in the +pGLO tube and mix the tube. We incubate the tubes on ice for 10 minutes. While the tubes are on ice, we label four agar plates. Then we heat shock the +pGLO and -pGLO tubes. After we open each tube and add 250 microliters of LB nutrient broth to the tubes and close it. We again incubate the tubes for 10 minutes. After we mix the cubes again and pipet 100 microliters of the transformation and control suspensions onto the appropriate plates. Using sterile loops we spread the suspensions evenly on the surfaces of the plates. Final we stack the plates and tape them together and place the stacks upside down in the 37 degrees Celsius incubator for one day. Then we clean up.
d) Hypothesis- I hypothesize that the transformed bacteria will glow green under ultraviolet light.
Procedure
Results
Discussion
a) Background- In this lab, we will be using a process known as genetic transformation, where we cause a change in an organism by inserting a gene into an organism. Genetic transformation has been used in many sectors of biotechnology. In farming, it has been used to foster resistance to frost, spoilage, and pests. In bioremediation, bacteria can be transformed to digest oil, which is useful in the cleanup of oil spills. In medicine, diseases caused by malfunctioning genes are now being able to be transformed in gene therapy. This could potentially mean that a person's cells that are making them sick could be transformed with healthy copies.
b) Purpose- The purpose of this lab is to show how by inserting a gene into an organism, we can cause a change in that organism. In this instance, we would be transforming a bacteria with a gene that codes for Green Fluorescent Protein that will cause the bacteria to fluoresce under ultraviolet light.
c) Process- First we are labeling one closed test tube +pGLO and another-pGLO. Then we pipet 250 microliters of transformation solution into the tubs and place them on ice. Using a loop, we pick up the +pGLO immerse the solution at the bottom of the tube and then do the same with -pGLO. Then we add 10 microliters of plasmid DNA directly to the liquid in the +pGLO tube and mix the tube. We incubate the tubes on ice for 10 minutes. While the tubes are on ice, we label four agar plates. Then we heat shock the +pGLO and -pGLO tubes. After we open each tube and add 250 microliters of LB nutrient broth to the tubes and close it. We again incubate the tubes for 10 minutes. After we mix the cubes again and pipet 100 microliters of the transformation and control suspensions onto the appropriate plates. Using sterile loops we spread the suspensions evenly on the surfaces of the plates. Final we stack the plates and tape them together and place the stacks upside down in the 37 degrees Celsius incubator for one day. Then we clean up.
d) Hypothesis- I hypothesize that the transformed bacteria will glow green under ultraviolet light.
Procedure
Results
Discussion
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