Disease Gene Lab Intro

Kary Mullis at Cetus Corporation created Polymerase Chain reaction in 1983. PCR is used to create a large amount of DNA in a test tube in order to be able to use it to identify specific genes and spots of interests. Any DNA sample can be used and mass produced such as a drop of blood or hair follicle. DNA  can be used to identify crime victoms, medical diagnosis etc. In this lab we will start on day one by taking DNA from our body such as a cheek swab after rinsing with a salty solution that is than transferred to a test tube. The test tube shall be heated up to over 95 degrees and instagene matrix shall be added so that DNase shall not break up the DNA that we want. This happens for 5 minutes after an incubation period of 10 minutes at 56 degrees celsius.
Next Polymerase Chain Reaction is used to amplify DNA by started with a template, mixing in individual deoxynucleotides, DNA polymerase, Magnesium ions as a catalyst, ogligonucleotide primers, and salt buffers. Copies are made doubling each cycle until there are millions of copies of the same strand of DNA. PCR begins in the denaturation step in which the mixture is heated to 94 degrees celsius for 1 minute seperated the double strand. Next the annealing step has oligonucleotide prmers attach to the complementary strands and ast as primers for polymerae making new complementary strands at 60 degrees celsius. Next the extension step adds the nucleotides to the primer actually making the complementary strand at 72 degrees celsius. These steps form one cycle usually has 40 cycles.
There are 23 pairs of chromosomes in the human genome with about 30k to 50k genes. The other 95 percent is noncoding DNA. In this activity we are lokking for the Alu element in the PV92 intron region. On the gel if both the inserts move at the same speed then they are both either dominant and will show by being slower, or recessive and be the same but moved farther up. If they are one of each then they are a mixture in which in theory if it was a disease the dominate would over ride and they would be a carrier.

GMO Results

In this lab we found that both the orange and the corn samples that we took were genetically modified. This makes sense because the orange that Daniel had was a cutie and corn is almost certainly genetically modified unless otherwise certified within the United States. The controls that were in place thankfully did not need to be used because the lab went splendid. The gel electrophoresis illustrated that the samples with added plant primers went farther and that those with GMO primers all matched up to each other. If they had not shown up in spots two and four we would assume that the samples were not genetically modified but this was not the case. Some sources of error may have been using a pestle and mortar instead of a blender provided us with a lesser amount of sample that then showed up very faintly within the gel. This was not too big of a deal because our results were spot on.

Experimentation Proving Most American Foods are Genetically Modified.

Introduction:

Genetically modified food contains genes implanted from other organisms for a specific purpose. They can be beneficial by providing more nutrition or yield, but the full effects are still unknown. In the US genetically modified foods do not have to be labelled as such which is potentially dangerous for those with allergies, and the public is unable to see the actual pros and cons of the food. Some cons include the potential for creation of superweeds by overuse of cross-pollination with herbicide resistant crops, and the creation of monocultures provides the potential for a single disease to wipe out main food crops such as corn. Also allergies are much more prone by using genes from different species. Some other pros are more storable foods(square), last longer, taste better, be more nutritious, can grow using lesss resources. In this lab we will be testing a variety of foods to determine there upbringing. We could specifically test individually the GMO proteins by useing Enzyme linked immunosorbent assay but unfortunately the genes are different in each organism. Instead we will a Polymerase Chain Reaction to identify the sequences inserted into the GM plant. This test can detect 85 percent of all GM crops because we are not looking for the gene but the plasmid that is transferred from the bacteria to the plant cell.

Procedure:In this lab we will start out by preparing the food and cutting a slice out around all the edges. We will hurt the cell wall and then open up the cell membrane. DNAse will be killed off with instagene maxtrix beads. On day two we wll do the polymerase chain reactions and amplify the DNA. Then the primers, DNA polymerase, nucleotide bases and reaction buffer will be mixed in a single test tube. Then the mixture will be placed in the thermal cycler, where it will cycle from 94 degrees Celsius to 59 degrees Celsius back and forth. The first step is called denaturation which happens at high temperatures. The next is the annealing step in which two template strands will compete with the primers but lose at low temperatures. Lastly the extension step will stay at 72 degrees Celsisus and the DNA polymerase will make copies of the DNA strand. It takes three full cycles that the fragments of precise length are generated. On the third day we will use gel electrophoresis to test with a GMO primer and have a plant primer set as a control.

Playing God; When Do I Learn How to Smite?

In this lab we will perform genetic transformation. Genetically Modified crops are made this way, insulin is created in bacteria for diabetes, and to digest oil spills etc. In this experiment we will be transform bacteria with a gene coding for Green Fluorescent Protein. Therefore the bacteria will glow green under ultraviolet light. In this experiment we will be transferring the gene from one organism to another with the help of a plasmid( contains DNA that bacteria keeps when it needs to adapt quickly.)  The pGLO plasmid contains penicillin resistance as well as the GFP, as well as a gene regulation system to control expression of the protein in transformed cells.
In this lab we will have three main steps to move the plasmid through the ecoli cell membrane
1.Usae a transformation solution of CaCl2.
2. Carry out heat shock.
growing the cells in presence of penicillin
3.Provide the cells with nutrients and an incubation period to allow them to express their new genes.

Microarraynalysis Results

In this experiment we found that there were two genes only turned on in cancerous cells(1 and 5) and two genes on in healthy cells (3 and 6). One of the other spots held a gene that was not turned on in either cell(4). The other was colored purple(2) and therefore on in both the cancerous and healthy tissues. The 4 differences of color between the two healthy blue genes and the red cancerous genes shows that the cancer cell is missing too necessary genes that probably kill unhealthy cells, and have two different cells turned on that probably send signals for the cancerous cell to continue to multiply. Sources of error could conclude from the short amount of time number 2 stayed purple or from the dye not being in high enough concentration. In conclusion this lab went very well.

Microarraynalysis Healthy and Cancerous Genes

In microarray analysis genes of comparing factors such as cancerous and healthy DNA are put onto DNA chip. The DNA chip would be used to analyze complementary DNA made from mRNA isolaed from cancerous and noncancerous tissue. The samples are flagged with dyes and the extent to which a flagged gene adheres to its complement reveals which genes show up in cancerous tissues as well as noncancerous tissues. This information is all processed by a computer and saves a lot of money and time. A single microarray can have more than 30,000 spots of DNA, each being a different gene. Some genes are not transcribed to RNA and eventually protein in cancerous cells and some are made too much of in cancerous cells. Finding the exact genes can help medical personal to create drugs to directly treat and eliminate cancer. We are personally using 6 genes in this lab for our microarray.