Gel electrophoresis

mL of 0.8% agarose gel is heated in the microwave for 1.5 minutes. The solution was cooled to room temperature and 5 μL of GelStarTM, a DNA marker, was added to the 50 mL. The DNA fragments will bind to GelStarTM and the GelStarTM enables the DNA complex to be visible under UV light. Carefully pour the solution into the gel-casting tray, insert the well comb and let the gel solidify. Place the gel tray so that it is parallel with the gel box and then fill the chamber of the gel box with 1XTBE…

The pre-cast gel was placed in a Pierce Precise Protein Gel Cassette. The gel was an 8x8 mini, 1mm thick and had 10 wells for loading samples. The inner chamber of the cassette was completely filled to the brim with 150 ml of Tris-Glycine gel running buffer, the mini tank outside was filled with the same buffer, to approximately 1/3 of the container. The protein samples were then…

Upon completion of the PCR, the final product should be highly concentrated DNA corresponding to the gene of interest. We ran our gene of interest on gel electrophoresis to compare our gene of interest to a marker of known size to confirm the size of our genomic DNA. On the image of the gel (figure 1), the sample containing the DNA ran, but the sample containing the DNA ladder did not. As a result, an appropriate determination of the sizes cannot be made because there is no ladder to compare it…

Introduction: Bacteria create an enormous domain of prokaryote microorganisms that are diverse and abundant in most habitats on Earth. In fact, bacterial cells drastically outnumber somatic cells just on the external part of the human body compared to somatic cells within the whole body of a single individual. Bacteria are found just about anywhere on earth and are usually alongside other microorganisms including: mold spores or yeast cells, creating some difficulty when trying to distinguish…

By Chelex based DNA isolation, DNA was isolated from buccal cells to study the genes TAS2R38, CDK3, ADH/ALDH, and D1S80. It was hypothesized that after gel electrophoresis, the TAS2R38 DNA sample will be cleaved at two places, since by the taste test was positive for PAV. The CDK3 gene will display one or two bands and the ADH/ALDH was expected to form one or three bands based on homozygosity or heterozygosity for the gene. D1S80 will have repeated sequences from 14 to 72 repeats. After…

In this lab, many materials were used including, EcoRl restriction enzyme, DNA from the crime scene and subjects 1 through 5, centrifuge, water bath, DNA loading dye, Electrophoresis chamber, TAE buffer, and a fast blast stain. The EcoRl restriction enzyme is the enzyme that was used in order to cut the DNA at specific sites into many separate fragments. A centrifuge is a machine that uses a rotating container inside it in order to force all of the liquids to the bottom of the test tubes within…

Report 5 Title Detecting possibility of expressing disease condition and presence of cystic fibrosis (CF) associated mutation (△F508 and G551D) in DNA sample using allele specific polymerase chain reaction (ASPCR). Abstract Elevated levels of blood immunoreactive trypsinogen (IRT) may be obtained in patients with cystic fibrosis (CF), when it is detected in primary screen, DNA analysis is performed to look for the 12 most common mutations associated with CF. This study aimed to investigate…

Gel electrophoresis allows the products of the PCR reactions to be analyzed in order to determine the identity of the tetracycline resistance plasmid. First, in order to see the DNA fragments, move across the gel, add 2 uL of bromopenol blue tracking dye to each of the tubes. In the first well add 10 uL of the PCR DNA ladder as a comparison. Carefully load 15 uL from each of the samples into the wells following the chart below. After the samples are loaded, place the lid onto the gel rig,…

Directions for the nano drop was run accordingly to machine directions. For electrophoresis, 10 µl of the sample was transferred into a clean microfuge tube with 2 µl of 6x loading dye. The portion of the sample was pipetted into an agarose gel and run at 120V for roughly 60minutes. Upon completion the gel was photographed for conformation. PCR Preparation A master mix was prepared in accordance to the PCR set up table provided by laboratory…

enable to do replication with PCR in both Watson(top) strand and Crick(bottom). The presence of Saw1 insertion gene will not matter in this assay. Therefore, the known and unknown will not show a difference in bands generated from the PCR and Gel electrophoresis. The second assay has PGEX-F1 primer and Saw1 primer. The Saw1 primer can either act as forward primer or reverse primer depends on the orientation of Saw1 gene insertion According to the manual, Saw1 acts as a forward primer for…