Rectal Cancer Case Study

The vital components and techniques of gene cloning are as follows, the DNA sequence that contains the desired gene (EZH2) is amplified by Polymerase chain reaction. PCR was established by Kary Mullis in 1985, popularly known to amplify target sequences of DNA (EZH2) to a billion fold in several hours using thermophilic polymerases (Taq) ,primers and other cofactors (Sambrook and Russell, 2001). Three crucial steps are involved which are Denaturation (at 95°), Annealing of the forward and reverse primers (55-65°) and lastly primer extension (at 72°). After amplification the desired sequence is integrated into the circular vector (pbluescript) forming the recombinant molecule. For the compatibility of the insert and vector, both were digested with (EcoR1) so the same cohesive ends are generated in both, making it easier to ligate. EcoR1 is a restriction enzyme that belongs to the type II endonuclease class which cuts within dsDNA at its recognition site “GAATTC” (Clark 2010; Sambrook and Russell, 2001).

There were several steps used to acquire the colony necessary for the PCR. First a student forearm was swabbed using a cotton swab, the cells were then placed in an agar plate. DNA was then extracted from the cultured bacteria by using a technique to lyse the cells and solubilize the DNA, then enzymes were used to remove contaminating proteins. The DNA extraction consisted of a lysis buffer that contained high concentrations of salt for denaturing. Binding with the use of ethanol and a washing step to purify the DNA. The final step for the DNA extraction was elution where the pure DNA was release. Proceeding the extraction of DNA the results of the 16s gene amplification were examined through gel electrophoresis it was analyzed by estimating the size of the PCR bands with marker bands. After measuring the success of the extraction, a technique called TA cloning was started. Cloning of PCR products was done by using partially purified amplified products with

The DNA extraction results, along with the PCR product, did not fare well. There was not enough product produced to be viable in the later stages of the experiment, so a backup was used in place of the original product.

The Basic Local Alignment Search Tool (BLAST) was the final portion of this lab report. BLAST is provided courtesy of the National Center for Biotechnology Information (NCBI). After running the five chosen tests, students were given the two 16S rRNA sequences that were associated with their assigned culture. The BLAST results served to confirm or disprove the hypothesis of what the unknown may be. The BLAST program compared the given 16S rRNA sequence to a database of known sequences and searched for similarity. (10) This search tool is capable of comparing nucleotide or protein sequences to find statistical significance between the matches. A perfect match to the unknown sequence is indicated by a

The present study targeted on methylation as a biomarker to show a particular epigenetic change. Methylation takes place when small chemical groups attach to the DNA, and it can impact how genes are converted into proteins.

Routine full blood count, liver function tests and urea and electrolytes were within normal range; however, she was found to have elevated carcinoembryonic antigen (CEA), a tumour marker associated with colorectal cancer. She was also positive for faecal occult blood. Subsequently, she underwent a colonoscopy with biopsy that confirmed the diagnosis of rectal adenocarcinoma. MRI staging was performed to determine the extent and spread of the disease that revealed to be Stage IIB on TNM

Colon cancer, also known as Colorectal cancer is cancer of the colon, which is the final part of your digestive system. Most of these cases begin as adenomatous polyps, which are small benign (noncancerous) cell clumps, but overtime these polyps can become cancerous.

The agreement of the MGMT gene methylation levels in the blood and rectal tissue was graded as good (κ=0.78) as 90% showed concordance and 10% showed discordance. While the agreement of the ERCC1 gene methylation was graded as very good (κ=0.85)

DNA was quantified and a luminometric methylation assay was performed. This assay is based upon

A person’s diet, as well as lack of physical activity and obese citizens is severely large risk factors too. Inflammatory bowel disease, which is often mistaken for colorectal cancer, too, plays a part in the risk factor. Smoking, which most people think would be the most common cause of lung cancer, has a higher colorectal cancer rate than the average person, as well as African American citizens. Colorectal cancer affects not only the colon itself, as well as a person’s genes. The APC, p53, and K-ras genes are commonly involved. The APC gene stands for the Adenomatous Polyposis Coli gene. It is known as a tumor suppressor gene, and mutations of it are found in common colon polyps or cancers. Furthermore, in the p53 gene, cells that have damaged DNA are repaired by this gene. As well as the APC gene, it is a tumor suppressor. When it’s mutated, it no longer functions, leaving damaged DNA cells in the body. Lastly, the K-ras gene helps with cellular growing and signaling. In the abnormal state, it can result in a continually growth-simulated state. The treatments for colorectal cancer may not help the mutation of these

Material and methods: 37 Egyptian patients diagnosed with rectal carcinoma were included in this retrospective study. 28 patients received neoadjuvant chemoradiation therapy while the remaining 9 cases did not. Total mesorectal excision was done by the same colorectal surgeon. The specimens of TME were examined to retrieve the lymph nodes.

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Colorectal cancer mainly starts at colon or the rectum. They are common in most of the way like features, but they have different treatment. What is the different between colon cancer and rectal cancer? Colon cancer happens first four to five feet of the large intestine and rectal cancer happens in the last few inches of the large intestine where it is connected to anus. (cancercenter.com)

Good quality genomic DNA was isolated (Sambrook, et al 2002) and bothansA1 and ansA3 genes were amplified by PCR. Clear bands of both the genes showing a size of 1kb were observed under UV transilluminator after agarose gel electrophoresis (Fig 1).

Also, DNA extraction, PCR and Illumina MiSeq sequencing were done on the soil, was noted that they knew to do this process since it was done on a by a previous study (Manli Wu, 2017). Multiple copies of the extracted 16S rRNA were done using 2 different primers and then sequenced on the Illumina MiSeq (Manli Wu, 2017).