DNA Fingerprinting Using Agarose Gel S. Aaron Sowards Bio 122 Lab 04 Brianna Adanitsch Jakob Lester Minhenga Ngijoi 2/21/18 Dr. Chad R. Sethman Abstract DNA fingerprinting is the process of analyzing an individual’s DNA base-pair patterns. The DNA fingerprinting lab involved identifying the suspect using Agarose Gel and Polymerase Chain Reaction. It was found that suspect two s DNA matched the crime scene DNA. This is known because suspect twos DNA traveled the same distance as the crime scene DNA. DNA Fingerprinting Using Agarose Gel Introduction In 1984 Dr. Alex Jeffreys came up with deoxyribonucleic acid (DNA) fingerprinting, which is also known as DNA profiling or DNA typing.
Next step is translation, in which the RNA becomes a protein, which then can act as structural units or enzymes. 2. How does DNA replicate itself? In order to replicate itself, DNA undergoes DNA replication, a process in which the DNA unwinds and splits in two. From that point on, new nucleotides are added to each of the original strands (A to T, C to G) until the result is two identical sequence copies of DNA.
After which the digestions were examined by gel electrophoresis. The samples were run on a 50 mL 0.9% (w/v) agarose gel in 1X TAE buffer at 100 V until the leading track dye traveled 2/3 the distance of the gel. The gel was then soaked in GelRed for 20 minutes and examined under UV light. To prepare the digestions 10 μL of each digestion was mixed with 2 μL of 6X track dye in a micro centrifuge tube. 12 μL of 1 kb DNA ladder and each digestion was run on the
Title: Genetic pGLO Transformation Introduction: Genetic transformation is a transformation that involves a change in genes. Transformation is the process by which the genetic material carried by a cell is altered by the incorporation of foreign exogenous DNA into its genome. In this lab, we used a procedure called heat shock, accompanied by a bacterial plasmid vector, to transform bacteria with a gene that codes for GFP (Green Fluorescent Protein). A vector is an agent “employed to transfer the gene from one organism to another” (Lab manual). Two common vectors are phages and plasmids.
Figure 20 and 21 casting tray Figure 22 gel box After 1 hr., take out the gel from gel box carefully, place it into the machine, so that the DNA gel electrophoresis can be visualize under UV light. Figure 23 gel documentation system, used to visualize gel electrophoresis with UV light NMR spectroscopy First of all, 3 samples were prepared, peptide in SDS, DPC, and buffer. The sample temperature was maintained at 298 K, prepared by supervisor and H(hydrogen) in SDS and DPC micelle was replaced with D(deuterium) , so that in proton NMR, peptide won’t be interfered by H in micelle. amount of peptide in sds and dpc In the case of NMR of peptide alone, sample was prepared with 90% of H2O which is 540μL and 10% of D2O which is 60μL, so 600μL of solution was used to dilute 1.5mg of peptide. Put 600μL of sample into NMR Sample Tubes, put the sample tube into NMR sample holder, and then run the test, the chemical shift of proton in peptide can be monitored.
However, when these bacteria are grouped together to have high cell density, the molecules they secrete amount to a certain number, and once that number is reached, the behavior of that bacteria is switched on and in this case, bioluminescence is created. Similarly, in my project, I am screening the anti bacterial activity using oils. Before I use the oil, I have to culture the bacteria overnight so that I could use them in the plates after 16-18 hours of incubation. Based on the talk, I believe that I have an idea on how bacteria grow. This Ted Talk has inspired me about science in numerous aspects.
However, all proteins are constructed from the same set of 20 amino acids linked in unbranched polymers. The covalent bond that exists between amino acids is called peptide bond, hence a polymer of amino acids is named polypeptide. A protein is a biological functional molecule made up of one or more polypeptides which is folded and coiled into unique three-dimensional structure. In laboratory, it is important to measure the concentration of proteins for research investigations. Biuret test is adopted to quantify proteins in fluid by using a spectrophotometer.
Amir Ahemedin Ms.Buckley Genetics 11/06/15 Transformation of E.coli Lab Purpose The purpose of this lab is to genetically engineer the E.coli strain by introducing two genes, the green fluorescent protein gene (GFP) and the ampicillin resistant gene (AMP). Then observe whether or not the E.Coli strain would take up these genes and become fluorescent. Background Information In this lab, bacterial transformation was one of the three processes that occurred when genetic material is introduced to a bacterial cell. Bacterial transformation is important because it allows for the cloning and movement of DNA between strains. This transformation usually occurs within plasmids, which are closed circular molecules made up of double stranded DNA.
The last error that may have occurred due to tape. Though caution was emphasized, and extreme carefulness had been placed in the taping process, some tape may have gotten inside the bottle, and fruit flies may have been stuck to it, which had skewed data. The lab manual from College Board, which states, “The common fruit fly….feeds on the fungi of rotting fruit,” (College Board 2004) reinforces the hypothesis clearly, and depicts chi square values with values such as 18.75. Experiments performed in UC Davis show that “fruit flies are regularly trapped….by yeast, and “the chemicals released by microbes…. [attracted] more flies.” (Garvey 2014) Some ways to improve the lab are to make sure that the error sources are fixed.
The STR length contrast is what is used to differentiate individuals. Gel electrophoresis then uses the STRs to create a DNA profile. The gel electrophoresis separates the STRs depending on their length and the pattern is then shown in fluorescent gel creating the profile. These profiles are then used by scientist to compare patterns between evidence and or suspects to determine a match or not a match. The probability of two people having the same amount of repeated sequences in STRs is one in billions of
The Solid sequencing platform, produced by Technologies/Applied Biosystems (ABI), performs sequencing by ligation method. Similar like the Roche 454 library preparation, genomic double strand DNA were sheared into small pieces and ligated with two types of adatptors P1 and P2 on two ends. One end with P1 adaptor binds onto the surface of the magnetic bead and emulsion PCR takes place to amplify single nucleotide fragment. Then the oil was washed out and four fluorescent labeled di-bases probes were added into the beads mixture. By matching the 1st and 2nd position of the template by di-base probes, fluorescence was detected and the extra tail with fluorescent probe is cleaved out.