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Student practical (3 activities in one ). Activity 1: Semi-quantitative gene expression by reverse transcriptase- PCR (RT-PCR) analysis. Activity 2: Extract total RNA from mammal tissues. Activity 3: gels result - Lab Report Example

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To study the gene expression of cyclin D1 in the given breast tumour and normal tissue specimens at mRNA level using semi-quantitative reverse transcriptase PCR and agarose gel electrophoresis techniques. To determine the relative amount of cyclin D1 mRNA present in the given…
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Student practical report (3 activities in one report). Activity 1: Semi-quantitative gene expression by reverse transcriptase- PCR (RT-PCR) analysis. Activity 2: Extract total RNA from mammal tissues. Activity 3: gels result
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Download file to see previous pages Alterations in the machinery that governs the progression from resting state into the cell cycle (the so-called G0 G1 transition) or from G1 into S phase are common to all tumor cells (Deshpande et al., 2005). It is the pRb pathway, which is the core decision-making machinery for progression from G1 to S phase that is commonly altered in several tumors. Metcalf (2010) described that cyclins are an evolutionarily conserved family of proteins that play an important role in the regulation of the cell cycle by binding to cyclin-dependent kinases (cdk). Normally, they are synthesized and destroyed in a precise manner and this periodicity enables specific cyclin-dependent kinase–cyclin complexes to facilitate the sequential events that take place during cell-cycle progression (Malumbres et al., 2009; Metcalf et al., 2010). The functions of cdk4 and cdk6, which are the partners of the D-type cyclins are dysregulated, which leads to an increased cell proliferation and turnover in mammalian cells (Metcalf et al., 2010). The D-type cyclins (D1, D2 and D3) are structurally and functionally similar proteins that are synthesized in response to mitogenic signals, bind to and form active complexes with cdk4 or cdk6, leading to phosphorylation and inactivation of pRb (Malumbres et al., 2009; Malumbres et al., 2004; Sherr, 1994; Metcalf et al., 2010). This in turn dissolves complexes of pRb with members of E2F family of transcription factors and associated chromatin-modifying enzymes, allowing transcription of genes required for S phase (Deshpande et al., 2005). According to a review by Deshpande et al. (2005), molecular analyses reveal the amplification, rearrangement of the cyclin D1 gene (localized on chromosome 11q13) as well as overexpression of cyclin D1 protein. It has been described in a wide spectrum of human cancers such as squamous cell carcinomas of head and neck, esophagus, tongue and larynx, carcinomas of uterine cervix, astrocytomas, non-small-cell lung ...Download file to see next pagesRead More
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