Gel Electrophoresis

Laura G onlyagher Partner Rob Einersen Biology Period D Mr. Alv atomic number 18z 15 February 2013 Gel Electrophoresis Introduction Agarose Gel Electrophoresis is a dish out in which the process of determining whether a strand of deoxyribonucleic acid is all cocksurely or prejudicially turn ond. The container in which the changeatin is stored has a shun and positive situation whichever side the desoxyribonucleic acid corpuscles go to means the desoxyribonucleic acid is charged the opposite way. (Ware, Lunte, Gardiner)For example if a DNA molecule goes to the negative side that means that the DNA is positively charged and vice versa.The agarose gel is composed of a fine powder substance, water and a airplane pilot solution. The solution must be boiled to its boiling point then you acquit to pour the solution into a casting m honest-to-god at that rest home also needs to be a comb that leaves six holds in the mold. It must be left in the casting mold for roughly a half an hour for the gel to solidify. Electrophoresis has a hardly a(prenominal) different uses such as establishing the size of a strand or molecule of DNA or RNA. (Bowen) It can also be use to find out family members or criminals or tests of that manner. This is all possible because of the DNAs charge and the charge of the molds.Although if a DNA molecule is overly big it is not firing to be able travel betting through the so it is better for the experimentation if the DNA molecules are elegant. To study the DNA small restriction enzymes are used. (Roberts) Restriction enzymes cut unique(predicate) DNA molecules in half which helps with the travel through the gel. Before the cataphoresis machine was developed people used to use gravity to set out DNADNA molecules are going to be negative and small molecules are going to move uttermostther than the larger molecules. The mapping of this lab is to learn how to bring to pass an agarose gel and properly load a well in an agar ose gel.The purpose is also to learning how to use electrophoresis equipment and how to analyze the results of DNA electrophoresis. Methods and Materials take leave off, by gathering all of the necessary materials for the beginning which is 1. 2 mL of gel mince in adept graduated cylinder, in a sepa identify graduated cylinder58. 8 mL of water and 0. 48 g of agarose powder. Next take an Erlenmeyer flask and mix all three of the substances, bulge out the flask on a hot shield and keep swooshing the mixture in the flask. Repeat that step until the liquid in the hot plate becomes clear, make sure that the solution does not boil.Let the solution sit until it is lukewarm. Now, the edge tray should be prepared by taping off the sides with painters tape so no liquid can escape. Place the comb in the designated slot in the molding tray and pour the agarose solution into the mold tray. Now, take a graduated cylinder and pour in 8 mL of the buffer solution and 400 mL of water, then, p our it into the gel box. After the gel solidifies, with extreme caution, remove the painters tape and comb. Next, take the mold tray and place it on the table now, take six disposable pipettes and the colored dye from you instructor and fill separately pipette with a different color.After each pipette has a different color, empty the pipette not too far in or too close to the surface of each well. Next, place the mold tray in the gel box, close the lid, and plug it in for just about a half an hour. After the half an hour is up, take a couple pictures of the mold tray and the results that you saw. Results Figure 1 Figure 2 As you see in figure 1, the dye pigments have been placed in each well and it is not perfect some of the dye have gotten out and leaked out on the top of the gel.That is okay though because the experiment was still a success because the dye pigments did move with the DNA molecules as you can see in figure 2. The dye pigments in all of the wells except for the w ell at the very top, all go towards the positive side of the tray. The well at the very top contained positively charged DNA so that DNA started to move towards the negative side of the tray. Discussion For the most part, my hypothesis was correct, I was correct about the smaller particles moving nevertheless than the larger particles. I was correct and incorrect when I said that The DNA molecules are going to be negative. I as correct because five out of six were negative, although at that place was one that was positive so it did not go in the same direction as the other five. DNAs direction is only influenced by one factor, which is whether the DNA is negatively or positively charged. This directly affects why the DNA moved the positive pole because DNA is negative so referable to attraction it goes to the positive side of the pole. When a molecule is moving its rate of length and speed correlates with its size, if a molecule of DNA is large, it is going to be harder for it t o move so it would be much easier for a small molecule of DNA to move across the gel.You can the molecules of DNA moving because they are dyed with the dye pigments. Electrophoresis causes the smaller DNA molecules to move further because it is easier for the positive charge to pull smaller pieces of DNA. Though, for this to happen, the power has to be morose on. Once the power is turned on it also turns on each ends charge on each side of the tray so the DNA is attracted to that side of the tray. Most of the DNA the molecules carried a negative charge. A negative charge is carried because those molecules went towards the positive pole.Although those few molecules carried a positive charge so they went to the negative pole. The banding patterns in the gel are opinionated by the size of the DNA molecule. It can be interpreted as some of the DNA molecules werent broken down as small as others. Scientists use the number of nucleotides in one sample and it is compared to another(preno minal) blood sample. All of the alikeities and differences add up. Also the sequences of the bases in a mountain chain of DNA. In many murder investigations, DNA is used to find the culprit. such as the outcome of James Anagnos, James was beaten and stabbed to death in 1977 in his bar. NBC) James was holding a strand of hair that belonged to his murderer. trine decades later they compared the DNA is the strand of hair to a man named frump Wright, it was a match. It didnt serve any justice though, because Wright died in 2002. This is similar to the case of Priscilla Ann Blevins. Priscillas remains were found off an interstate and they were stored in a facility and kept at a lower place the name Jane Doe. (Lohr) Blevins genic information was entered into a computer and it matched up with Jane Doe and was later sustain with dental records.DNA helped to resolve this 37 year old cold case just like it did for the 1993 murder of Alie Berrelez. Alie was a five year old girl who wa s sitting in her apartment complex eating pizza pie when she was kidnapped. (Curry) Alies remains were found four days later stashed succeeding(a) to a creek. Nick Stofer was the main suspect but they couldnt assay him because they didnt have enough evidence against him. When this crime happened there was no such thing as DNA testing so there was no way they could prove it was him. In 2011, they compared DNA from Alies under to Nick Stofer and it was a match.Again Stofer died before he could stand trial, he died in 2001. Bibliography B. R. Ware,Susan Lunte,Kathleen Gardiner, 2012, Electrophoresis, in AccessScience, McGraw-Hill Education, Retrieved from http//www. accessscience. com/content. aspx? searchStr=Electrophoresis&id=226400 Bowen R. , 2000, Agarose Gel Electrophoresis of DNA, in Colostate, Retrieved from http//arbl. cvmbs. colostate. edu/hbooks/genetic science/biotech/gels/agardna. html Richard Roberts, 2012, Restriction enzyme, in AccessScience, McGraw-Hill Education, Re trieved from http//www. accessscience. com/content. aspx? earchStr=restriction+enzymes&id=584150 2010, Three Decade old Murder Mystery Solved using DNA , in NBC California, retrieved from http//www. nbclosangeles. com/ intelligence operation/local/Three-Decade-Old-Murder-Mystery-Solved-Using-DNA-101944298. html Lohr D. , After 37 Years, Priscilla Ann Blevins Disappearance Solved Using DNA, Huffington Post, Retrieved from http//www. huffingtonpost. com/2012/11/01/priscilla-ann-blevins_n_2059155. htmlCurry C. , 2011, Cold Case of Murdered 5-Year-Old Alie Berrelez Solved, first rudiment News, retrieved from http//abcnews. go. com/US/cold-case-year-murdered-1993-solved-dna/story? id=14510785