RNA polymerase

4. Multiplex Polymerase Chain Reaction Multiplex PCR is molecular biology technique for amplification of multiple targets in a single PCR reaction. Indeed, multiplex PCR is a modification of conventional or realtime PCR in which two or more different PCR products are amplified within a single reaction (Henegariu, 1997). In a multiplexing assay, more than one target sequence can be amplified using multiple primer pairs in a reaction mixture. As an extension to practical use of PCR, this technique…

DNA samples were extracted from sixteen unidentified shark tissue specimens and underwent PCR amplification of CO1 mitochondrial gene. DNA samples were assessed for presence and quality using agarose gel electrophoresis. Species identification was performed by importing finalized CO1 sequences into two DNA databases to ensure accurate identification. Phylogenic analysis was then conducted to determine relationships between identified species. Electrophoresis Electrophoresis of DNA run in 2%…

minute by placing an anode (positive poled) at the bottom and a cathode (negative poled) at the top. PCR clean-up was tested using the same electrophoresis method as the DNA extraction. PCR A portion of DNA from the extraction was amplified using polymerase chain reaction (PCR). A new sterile tube was used. In the tube 23.0µL of master mix was added which contained 10.5µL of water, 12.5µL Green Taq( premix with buffer, Taq, and dNTPs). Next 0.5µL…

I also prefer the term "melting", though I recognise that this term is potentially confusing for a lay or beginners' audience. The analogy with melting of water ice, while intuitive, may invite the misconception that DNA "dissolves" into very small molecules rather than separates into its two strands. The lede states "These thermal cycling steps are necessary to physically separate the strands (at high temperatures) in a DNA double helix (DNA melting)", and in the description of the PCR steps it…

molecular genetics and specifically forensic analysis [1]. The goal of this experiment is to observe the tandem repeats of the VNTR (D1S80) locus through molecular lab techniques. Cheek cells were isolated from an individual and amplified via a polymerase chain reaction (PCR). Then, the amplified sample was separated through gel electrophoresis. This project entails the interpretation of fluorescing bands from gel electrophoresis in order to determine the number of tandem repeats present on an…

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 during the finding of microbial communities. Another method that was used during the project was whole genome shotgun metagenomic sequencing. These methods were used to obtain more accurate data…

In 1859, Charles Darwin documented that evolution of life on Earth had taken place and proposed the idea that natural selection was to blame in his book, The Origin of Species (Leicht and McAllister, 2016). Darwin’s findings arose from the different heritable traits, which lead to differences among alleles in a population. Genetic variation is the differences in genetic frequencies in a population and is the basis of evolution. The Hardy Weinberg Principle was developed to test for genetic…

For each group cDNA was synthesized from 500 ng of RNA using Maxima Universal First Strand cDNA Synthesis kit (Thermo Scientific, catalogue no-K1661) and consecutively 50 ng of cDNA was used for quantitative real time PCR assay. cDNA was amplified in a thermal cycle (CFX96 PCR system, BioRadLaboretories)…

from one generation to the next and each allele frequency. This theory notifies us by how Mendelian inheritance was the lead for all genetic variation and it can be used to follow Mendelian’s rules of segregation and the combination of alleles. Polymerase chain reaction was one of the essentials that is used in the Hardy Weinberg theory of genetic equilibrium which was used in determining the presence of ALU element using TBA 25. The importance of using PCR is to make more copies of small pieces…

denaturing when the double-stranded DNA is separated into two strands. Second, annealing which enables the DNA primers to attach to the DNA when the temperature is lowered. Lastly, extending which is when a new strand of DNA is created by the Taq polymerase enzyme when the temperature is raised. This process is run in cycles 29 times and takes approximately 4 hours to complete. The PCR test was completed prior to the testing and was then given to each student to perform gel electrophoresis. The…