Once the gel hardened, .5X TBE (44.5 mM Tris base, 44.5 mM boric acid, and 1.0 mM EDTA) was added just until the gel was covered with the TBE buffer. Each sample was loaded into the gel as well as 10 μL of DNA size markers (1kb ladder, New England Biolabs) into a separate lane. The gel was allowed to harden at room temperature and then electrophoresed at 100 volts for 75 minutes. Using a UV imager, a photo was taken of the resulting traveled DNA fragments in the gel.
Next one microliter of DNA was mixed with one microliter of loading dye using a pipettor and loaded into the well. The same mixing process was completed for the PCR product, using one microliter of PRC product and one microliter of loading dye. For the purposes of this experiment, the DNA product was loaded into well six and the PCR product was loaded into well seven. Initially DNA was loaded into well five, however gel was pierced so samples were moved one well to the right. The gel was run at 100 V for one hour.
Both DNA and RNA has a maximum absorbance of 260 nm. The absorbance of 260/280 should be in between 1.8 and 1.9 to represent a pure sample of DNA. If the reading is higher than 1.9 then there is RNA contamination and if the reading is less than 1.8 there is protein contamination.
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.
It is vey important crime investigation feild where the criminal can traced out. It also plays a major role in the law of justice where people lie about their relation that they had with other people(father and son or mother and son). 5.Below are the images of the gels after electrophoresis. Indicate which lane is loaded with the samples from your group?
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.
It has been said to have revolutionized forensic investigations. Deoxyribonucleic acid profiling has been used to exonerate the wrongly accused, convict the correct criminal, and even do something as little as finding out who the father of a child is (Roewer, Lutz). Deoxyribonucleic acid today can be obtained from processing bodily materials such as spit, semen, or blood. Because of the fact that all cells in the human body contain a nucleus and nuclei contain DNA, virtually any evidence left behind that is a bodily fluid can be tested for Deoxyribonucleic acid to help tie or eliminate those from a crime
Discussion PV92 Gel Electrophoresis Results: Through the usage of gel electrophoresis the correct allele for each sample was able to be determined. Lanes one through three in the gel,were the positive control lanes they contained the PCR cocktail and a known high-quality template for the PCR reaction. First lane contained the sample with the +/+ allele, which had two copies of the ALU repeat allele. The first lane had a band at about 941 base pairs.
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.
1.1 Abstract The purpose of quantitative analysis of protein using a spectrophotometer is to measure the concentration of proteins in a given sample. The experiment is conducted by laboratory method (Biuret Test) and using spectrophotometer to analyze the absorbance of reactants at 540 nm, hence determining the concentration of the proteins in a given sample. The purpose of stopped enzyme assay to study B-galactosidase is to determine the effect of temperature and concentrations of substrate on enzyme activity.
The purpose of this lab is to determine the relationship that exists between the number of amylase gene copies and ancestral diet. As the human civilization moved forward toward agriculture the diets of humans also changed. Depending on where the humans originated would give insight to how much of their diet was starch based. My family’s geographic origins are from China. Thus knowing that the country has a high starch based diet, we would suggest that I would have a high amylase production.
The DNA gathered by the group bore positive results only on Test for Deoxyribose; compared to the standard solution, which bore positive results on all chemical tests, namely, Test for Deoxyribose, Test for Phosphate, Test for Purines, and test for Pyrimidines. Introduction Nucleic Acid is one of the essential biochemical molecules
The streaking technique used was a modified streaking for isolation with a heavy quadrant one. The result revealed that bacteria is alpha, with an incomplete breakdown of the medium with a susceptibility of 17mm from the bacitracin gamma hemolysis. That is why the organism represented by the bar graph was in low numbers because it was incomplete. The other test was DNase agar, it is an enzyme test used to identify if the organism has the enzyme DNA. The streaking technique is a single straight line down the middle of the plate.
In this three-week long experiment conducted in the Bio 13 Lab, we were able to analyze a single nucleotide polymorphism (SNP) in our own genomic DNA and then determine our genotype at this specific SNP. In week one, we extracted genomic DNA from our cheek cells with swabs and prepared our DNA for PCR (Polymerase Chain Reaction) that would amplify the region with the intended SNP of interest. After one week and after the PCR was run outside of the lab section, the resulting PCR product was purified and treated with restriction enzyme Ahdl in order to prepare for the final analysis of our genotypes. In the third and final week of the project, we analyzed our PCR products by means of agarose gel electrophoresis. By the conclusion of the experiment, we had completed the analysis at the SNP of interest and determined our genotypes for this SNP.