Dna Analysis Dna Fragmentation Essay - 2433 Words

DNA ladder assays were used to evaluate the DNA fragmentation triggered through apoptosis when thymocytes were exposed to various concentration of diethylstilbestrol (DES). There are numerous methods for the quantification of DNA fragmentation but for this study, a lysis and DMSO protocol were used for the detection of apoptosis. Apoptotic DNA fragmentation was analyzed using 2% agarose gels to reveal a ladder-like pattern within each sample. The present study demonstrates a comparison between two DNA fragmentation protocols, lysis and DMSO protocol, to demonstrate the effects Dexamethasome had on thymocytes DNA. The DMSO protocol is time efficient, involving the direct lysis of cells with dimethyl sulphoxide (DMSO), vortexing, adding of 2% SDS-TE buffer and centrifugation. While the lysis protocol is time consuming, involving the use of HBSS, the direct lysis of cells using a lysis buffer, incubation period, RNase A and centrifugation. Upon comparing both methods, the lysis protocol provided better results than the DMSO protocol because it produced a more defined bands. INTRODUCTION Immune System The immune system serves as the body’s defense mechanism against infectious invaders such as bacteria, viruses, parasites, and much more. The immune system has two types of immunities: innate and adaptive. Innate immunity refers to the defense system you are born with. It helps to protect you against antigens and has barriers that keep harmful materials from entering theShow MoreRelatedPerl And Bioinformatic Data1053 Words   |  5 PagesAn example of further investigations into analysis include functions such as randomization, which can be applied to emulate and investigate mechanisms of mutations in DNA and the effect they have upon biological activity of their associated counterparts. Additionally, Perl can handle many forms of data from numerous databases including FASTA style data, genBank, PDB and BLAST. In an ever-growing sea of bioinformatic data, Perl, among others has addressed a way of divulging, searching and storingRead MoreSite Directed Mutagenesis ( Sdm ) Technique942 Words   |  4 PagesSite-directed mutagenesis (SDM) technique is commonly used to induce desired change in DNA plasmid sequence by mutation, insertion or deletion with oligonucleotide primers (1). This SDM usually cooperate with ploymerase chain reaction (PCR) as to amplify the concentration of mutated template (2). PCR, a temperature-based cycle reaction, is completed with three initial steps including denaturing the DNA template, anneal the mutated oligonucleotide primers and elongating the mutated primer with ploymeraseRead MoreDna Digestion and Electrophoresis728 Words   |  3 PagesDNA DIGESTION AND ELECTROPHORESIS In this experiment we will be doing a process called as DNA digestion or also known as restriction digest. A  restriction digest  is a procedure used in  molecular biology  to prepare  DNA  for analysis or other processing. It is sometimes termed  DNA fragmentation, scientists Hartl and Jones describe it this way: This enzymatic technique can be used for cleaving DNA molecules at specific sites, ensuring that all DNA fragments that contain a particular sequence have theRead MoreAnticanbar Drugs1617 Words   |  7 Pagesscavenging capacity and an interaction with antioxidative enzyme utilities. Some treatments such as metformin and TQ as well as natural products from the crude black seed are examined as synthetic or natural antioxidants to avert oxidative damage of DNA caused by reactive oxygen species during cancer formation [30]. The present data have shown that the treatment with the IC50 concentration of metformin alone caused a significant increase in the activities of SOD, CAT and GSH levels while decreasedRead MoreOrigin Of Replication Is The Starting Point Of Dna Replication Essay1729 Words   |  7 Pages(Nov-9-2016) Origin of replication is the starting point of DNA replication. It cannot replicate in the host cells without this site. Antibiotic resistance gene can be served as a select agent to find the right clone and then improve the plasmid transformation efficiency. Antibiotic resistance gene can also give the bacteria a pressure to have the plasmid, because the bacteria would use more energy to replicate both the plasmid and its own DNA. (1) The protein needs complex eukaryotic post-translationalRead MoreWhat Protective Effect Of Α -lipoic Acid ( La ) Against Oxidative Stress And Apoptosis1696 Words   |  7 Pagesbiochemical and histopathological studies. Malondialdehyde (MDA), reduced glutathione (GSH) contents, catalase, superoxide dismutase activity (SOD), integrity of nuclear DNA (nDNA) were estimated. Immunostaining was used to determine Bax and PCNA localization. MTX-treated rats showed increased levels of MDA and fragmentation of DNA in addition to reduction of GSH levels and activities of catalase and SOD. Histological examination of MTX-treated rats demonstrated degenerative changes that involved theRead MoreWhat Is The Free Amino Groups Of Nucleobases?857 Words   |  4 Pagesgroups of nitrogen bases of nucleosides and nucleotides under alkaline pH conditions are also prone to the Maillard reaction (Reihl, O. et al, 2004). DNA reacts with reducing sugars (glucose, fructose, ribose etc.) and advances via Amadori reaction leading to the formation of irreversible complexes known as DNA-advanced glycation end products (DNA-AGEs) (Bidmon, C. et al 2007; Frischmann, M. et al 2005). Various report suggested that nucleobases possessing amine groups are susceptible to glycationRead MoreDiabetes Case Study1331 Words   |  6 PagesP. Talalay, Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors, Advances in Enzyme Regulation. 22 (1984) 27-55. [12] N. Shiraishi, Y. Inai, W. Bi, M. Nishikimi, Fragmentation and dimerization of copper-loaded prion protein by copper-catalysed oxidation, Biochem. J. 1:387 (Pt1) (2005) 247-55. [13] B.K. Sinha, M.A. Trush, K.A. Kennedt, E.G. Mimnaugh, Enzymatic activation and binding of adriamycin ton nuclear DNA, Cancer Res. 44 (1984) 2892Read MoreThe Human Genome and DNA Sequencing1335 Words   |  5 PagesA genome is the complete set of DNA, including all of its genes. Each genome contains all the information needed to build that organism. In humans, more than 3 billion DNA base pairs are present. For advance knowledge of molecular and evolutionary biology, it is crucial to sequence the DNA of every human chromosome. This is quite huge in scale, as it sought to determine the order of all 3 billion nucleotides in the human genome. Hence a number of sequencing techniques were developed that at the sameRead MoreThe Method Of Detection Of Dna1318 Words   |  6 Pages1. Primer Annealing – temperature is lowered so that primers attach to DNA strands 2. Extension – DNA polymerase (e.g. Taq) helps to make new DNA strands. The PCR cycle continues as DNA is copied exponentially. The original PCR methods allowed detection of DNA growth at the end of the process; however the introduction Real Time-PCR (RT-PCR) has allowed DNA amplification progress to be monitored throughout the assay (6). RT-PCR is often confused with reverse-transcriptase PCR (rt-PCR), however rt-PCR