Rapid Colony Transformation of E -Coli with Plasmid DNA Lab Report

Rapid Colony Transformation of E -Coli with Plasmid DNA - Lab Report Example The paper "Rapid Colony Transformation of E -Coli with Plasmid DNA" analyzes E-Coli transformed by plasmid DNA using a rapid method. The transformation method can be categorized into four stages. The first is Pre-incubation stage. The low temperature gels the cell membrane, thereby stabilizing the distribution of charged phosphates and allowing them a more effective shield from the cat ions. The second stage is Incubation. The DNA is added and the cell suspension is kept at 0Â ºC.the cat ions are thought to neutralize negatively charged phosphates in the DNA and the cell membrane. The third stage is Heat Shock. The cell +DNA suspension is briefly incubated at 42Â ºC and then returned to 0Â ºC. The rapid temperature change creates a heat imbalance on either side of the E-Coli membrane and is supposed to create a wave that sweeps plasmids into the cell. The fourth stage is Recovery. LB broth is added to the DNA/cell suspension and incubated at 37Â ºC before being put on plates with different selected antibiotic resistant markers. Transformed cells recover from the treatment, amplify the transformed plasmid and begin to express the antibiotic resistant strain. Samples of E-coli cells are taken from a nutrient agar plate (LB agar) and suspended in two tubes containing a solution of calcium chloride. Plasmid pAMP is added to one cell suspensionBothe the tubes are then incubated at 0Â ºC for 15 minutes. After this a brief heat shock is administered at 42Â º .the samples are cooled and LB broth is added.

The Financing of Small Businesses in Saudi Arabia Essay

The Financing of Small Businesses in Saudi Arabia - Essay Example Moreover, differences have been identified between the measures taken by governments in developed and developing countries regarding the limitation of inequalities in the support offered to entrepreneurs worldwide; the form and the level of governmental support to business activities have been also found to be related with the size of the firm involved – a fact that can offer the chance to foreign firms to enter the market – domestic firms that are already well developed are also likely to increase their power (Keren et al., 2002, 17). It seems that the financing of small firms can be a challenging effort in most markets internationally. Of course, the personal involvement of the entrepreneur – owner of a small firm – in the relevant effort could increase the chances for success – however, a positive outcome cannot be guaranteed. Various factors, like the local culture and ethics and the position/ strength of the national economy, seem to influence the effectiveness of the particular initiatives. The above issue is highlighted in the study of Busenitz et al (2001). The above study refers to the effectiveness of strategies of small firms in China; it has been proved that ‘younger business owners with a higher need for achievement, greater commitment, and perceived limitations in physical facilities were more likely to expand’ (Busenitz et al., 2001, 12). From a different point of view, it is supported that ‘differences in environmental conditions in different country markets, in terms, for example, of market size and growth, rate of technological change, or barriers to entry, may also lead to differences in strategy’ (Douglas et al., 1989, 440). The view of Douglas et al. (1989) highlights the potential involvement of a series of factors in the development of business activities – in accordance with this view, the local culture can affect business plans –almost at the same level with the size of the firm.