16S ribosomal RNA

To confirm the amplification of the 16S rRNA gene of the bacteria you use the performing gel electrophoresis. The PCR DNA is used to obtain a DNA sequence and use the DNA sequencing software to edit and use the NCBI database to find matching 16S rRNA gene sequence using BLASTn. There are many tests you can do to find our what your unknown bacteria are (Leicht, 2015). For this experiment my partner and I selected bacteria that was on our finger. We thumb printed the petri dish and let the bacteria grow on it. We did a visual classification of the DNA. We determined our bacteria were gram positive after doing the KOH string test. Then we preform additional testing using different growth mediums like vancomyin, EMB-lactose, PEA, and MSA, along with a catalase test. We also amplified the 16S rRNA gene from the unknown bacterial DNA used gel electrophoresis used Finch TV sequence to determine what the unknown bacteria is (Leicht, 2015). After preforming several tests that were expressed above, we ended up distinguishing that our unknown bacterium is Lactobacillus jensenii 269-3 (Altchul, 1990). Materials and Methods: My group and I obtain some bacteria from our fingerprints and different locations around the Biology Building. My partner and I examined our bacterial colonies that grew from the cells collected. We selected a single colony of the unknown bacteria from a fingerprint. We set up a PCR reaction to amplify the 16S ribosomal RNA gene, of the bacterial genome. To do…

A genotypic method uses molecular techniques to identify bacteria by doing DNA or RNA analysis of the bacterium’s genome ("Conventional Bacterial Identification Methods", 2013). The 16S rRNA gene sequences provides species-specific signature sequences useful for bacterial identification ("16S Ribosomal RNA Sequencing Theory”, 2016). The 16s rRNA gene sequence uses a polymerase chain reaction (PCR) to amplify a specific DNA target from a mixture of DNA molecules, which helps in the…

Some of the features that would be used for analysis were population, genotype, disease, nutrition, medication as well as environment and influence on disease. The key findings of the project would later be placed in a database that would be available to the scientific community. Hence sometimes the human microbiome project may be called a community project. One of the key methods used in the project was the targeting of 16s ribosomal RNA gene sequences these were used as taxonomic makers…

PCR is a common practice used to amplify a certain region of DNA. In this experiment, PCR was used to amplify the 16S ribosomal RNA gene of the bacteria. This locus was used because it is one that evolves slowly in bacteria. A Taq polymerase is used for this PCR because of the temperatures the reaction goes through. The16S rRNA primers used in this PCR, are complimentary to sequences conserved in all bacteria and will target sites of variation. These sites of variation were the regions of the…

can provide a more accurate picture of the diversity and structure of the whole microbial community. For bioturbation studies in particular, techniques such as denaturing gradient gel electrophoresis (DGGE), restriction fragment length polymorphism (RFLP), and ribosomal intergenic spacer analysis (RISA) have been used. Generally, diverse communities have been identified in burrow walls, and found to be more similar to either surface communities (Laverock et al., 2010, Pischedda et al., 2011) or…

transcriptional machinery of RNA Polymerase I in the oocyte Experimental Approach: Aim1: To address the roles of the Hira complex in rRNA transcription and embryonic development The Hira histone chaperone complex is an evolutionarily conserved complex, which is composed of Hira, Ubn1 and Cabin1, and cooperates with the histone chaperone Asf1a to mediate H3.3-specific binding and chromatin deposition12,13. I previously demonstrated the critical roles of maternal Hira /H3.3 on paternal…

Alkaline phosphatase (AP) is a homodimeric enzyme complex that is commonly found in a wide range of organisms, from bacteria to all tissues of the human body. AP is a zinc metalloenzyme (1), in which metal ions play a key role in the regulation of catalytic activities and stabilization of enzyme-substrate complex. As proposed by Gettins and Coleman using NMR studies (10), each active site of AP comprises of three metal binding sites, which acknowledged as M1, M2, and M3. Two zinc ions bind to…

so much behind the buildup of protein sequences. By the end of 2007, there have been forty four,272 protein structures deposited within the protein data Bank (PDB) SECONDARY STRUCTURE: Secondary structure is that the native arrangement of a polypeptide’s backbone atoms while not relevance the conformations of its aspect chains.It is a group of techniques in bioinformatics to predict the native secondary structures of proteins and RNA sequences based mostly solely on data of…

cancer has been made evident the use in studying this circadian clock gene is quite clear and the need for a more solid grasp on its mechanism is necessary to understand a possible treatment in cancer. For this experiment we chose a knockdown experiment through RNA interference (RNAi) of the timeless gene in order to observe the phenotypic outcome in planarian when this gene is absent. This is a known procedure for disturbing the function of genes in planarian (4). We made a double stranded…

2-cyanoethyl carbonochloridate (S.27b ) or 1-((2-cyanoethoxy)carbonyl)-3-methyl-1H -imidazolium chloride (S.27c) (Figure X.X.X), which were synthesized by the modified procedures of Merk et al. (2000) and Wielser et al. (1996). Deproetection was accomplished under nonprotic conditions using non-nucleophilic strong base 1,8-diazabicylco [5.4.0]undec-7-ene (DBU). The mechanism of deprotection follows -elimination pathway. Another advantage of this protection strategy is that the deprotection of…