Southern Blotting

(PCR), which isolates small fragments of DNA that have a high degree of variability from

Polymerase Chain Reaction was run and the results were placed into an electrophoresis gel to visualize the PCR product. To determine the alleles in the gene, a digest enzyme, Fnu4HI, was placed into the DNA. Tasters would view a cleavage formed at a restriction site in at least one of the alleles, whereas non-tasters would not.

This however, was unsuccessful, as the restriction digestion created fragments of many different lengths, resulting in smears at different lengths on the gel electrophoresis due to the restriction enzyme recognition sites being present in-between the telomeric repeat sequences, the analysis was thus

Figure 1 Gel Electrophoresis for Replication Taster PTC. The gel is composed of an ethidium bromide stained 3% agarose gel demonstrating DNA fragments which were a depiction of PCR amplification. The agarose gel contains nine loading samples, including from left to right, the MW marker lane 1 precision mol mass standard, lane 2 TB undigested PTC (5µl of DNA, 5µl of master mix P, and 2.5µl of loading dye), lane 3 TB digested PTC (5µl of DNA, 5µl of master mix P, 2µl Fnu4HI, and 3µl of loading dye), lane 4 TB A(L)DH G (10µl DNA, 10µl master mix G, and 5µl loading dye), lane 5 TB A(L)DH A (10µl DNA, 10µl master mix A, and 5µl loading dye), lane 6 MG undigested PTC (5µl of DNA, 5µl of master mix P, and 2.5µl of loading dye), lane 7 MG digested PTC (5µl of DNA, 5µl of master mix P, 2µl Fnu4HI, and 3µl of loading dye), lane 8 MG A(L)DH G (10µl DNA, 10µl master mix G, and 5µl loading dye), lane 9 MG A(L)DH A (10µl DNA, 10µl master mix A, and 5µl loading dye).

Gel electrophoresis is a procedure used in laboratories to separate DNA, as well as RNA and proteins. A gel slab is placed in a buffer-filled box and an electrical field is applied. The negatively charged DNA will migrate towards the positively charged side, where it can then be recorded and further analyzed.

Restriction enzymes cut DNA at certain sites to create multiple DNA fragments. Restriction enzyme HindIII has known DNA fragment lengths and recognition sites when digesting lambda DNA, while the lambda DNA recognition site for restriction enzyme XhoI is unknown. The goal of this study is to determine the lambda recognition site of XhoI by comparing a HindIII digest and a HindIII and XhoI double digest on an electrophoresis gel. The HindIII digest had a band at 9.4 kb, but this band was not visible in the double digest, therefore we concluded the recognition site for XhoI was around 9.4kb. There were also two additional DNA

After completion of the restriction digest analysis, the DNA could be distinguished from one another as completion of agarose gel electrophoresis allows for the different lengths of the DNA to be observed, as GST would be shorter due to having less base pairs than GST-Cherry. Another indicator was the use of the agar plates in determining the type of DNA. In our case, our DNA had more of a clear cloudy appearance, while some others had more of a purple colored DNA. Due to this indication, it was confirmed that the different types of plasmid DNA could be differentiated based on the color of the bacteria on the agar plate. Despite the use of gel electrophoresis, our results didn’t turn out very well and weren’t very conclusive in determining the actual identity of our bacteria. It would be advantageous to retry the gel electrophoresis in order to further explore the identity of the DNA. Yet, the agar plates led to us determining what the actual identity of our sample was. Thus, in conclusion, our bacterial sample was normal GST because both of our plates, #16 and #23, contained a clear cloudy bacteria that wasn’t colored purple in comparison to the GST-cherry

To decipher if a species by its morphology can be suggested as a hypothesis, but the results of its DNA will identify the species accurately. Tissues samples can be taken from the species in question, and the DNA can be extracted from tissue. Once the DNA is extracted it can be amplified. DNA can be amplified by the PCR procedure, in which specific gene regions can be used as barcodes to identify the species. These specific regions are known as Cytochrome oxidase 1 and Cytochrome B.

In this experiment, host NM554, a particular strain of E. coli, was used to cultivate human genes (Dolf, 2013). Through the use of cosmids, plasmids that carry the cos gene, DNA fragments were introduced into the E. coli and packaged into phage particles (McClean, 1998). Pst I is a restriction endonuclease, an enzyme that cuts DNA at restriction sites (Restriction endonuclease). The Pst I digest of human DNA in this study produced the DNA fragments that were examined. The dideoxynucleotide chain-reaction procedure, also known as Sanger sequencing, is the process of lengthening DNA using DNA polymerase to add on deoxynucleotides until a dideoxynucleotide is added on randomly (Rogers). Fluorescence in situ hybridization (represented by the acronym

These strands are separated by length using electrophoresis and detected automatically by computers to be analyzed (Lyons, 2004). Another method of genetic testing is extracting one cell from an 8-cell embryo and using preimplantation genetic diagnosis (PDG) to determine the presence of disorder-causing genes (Holt, 2012).

Western Blotting can be used to detect the Myosin actin light chain in different species of fish and is used to distinguish from different species based on variation, commonality, or evolutionary divergence. First, proteins are extracted from the tissue and loaded into a gel matrix. The matrix will separate the proteins according to size using an electric current. Proteins that are separated after are blotted from the gel and onto a paper membrane. An antibody is then added to the membrane paper and causes a colored reaction. Following the reaction, the results

The DNA sample to be sequenced is combined with a primer, DNA polymerase, and DNA nucleotides (dATP, dTTP, dGTP, and dCTP). The four dye-labeled,

The polymerase chain reaction or PCR for short can be used to create many copies of DNA. This allows the DNA to then be visualized using a dye like ethidium bromide after gel electrophoresis. The process has been refined over the years, however the basic steps are similar.

Gel electrophoresis is a method of taking DNA samples and turning them into visuals that can be compared and analyzed. First, DNA fragments are placed on a layer of agarose gel, and when electricity is added, the DNA fragments move through the gel. The smaller fragments move faster than the larger ones. Afterward, you can see how the different lengths of

Restriction Enzyme Digestion – The experiment was begun after putting on gloves to avoid any chemical contact with the skin. Four microtest tubes were obtained, and each of them was labeled to contain the different enzymes or suspect DNA. Two of the microtest tubes were used for suspect one and the two different restriction enzymes, while two other microtest tubes were labeled for suspect two and the two restriction enzymes. After labeling the tubes, the contents that were at the bottom were taken out by slightly tapping them. Then to begin setting up the enzyme reactions, a micropipette was used to obtain 10 μL of the reaction buffer which was added to each of the four test tubes. The buffer is important because it carries the electrical current from the power supply in the gel. After the reaction buffer was in each, the microtest tubes were individually filled with their specific enzymes and DNA, shown in summary through Table 1.1 below. The restriction enzymes are used to cleave the DNA at specific