@@ -272,15 +272,15 @@ export function Engineering() {
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The protocol entailed the utilization of varying concentrations of Lipofectamine 3000, specifically 1 µl and 1.5 µl, with a DNA quantity of 1 µg or 0.5 µg.
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The protocol entailed the utilization of varying concentrations of Lipofectamine 3000, specifically 1 µl and 1.5 µl, with a DNA quantity of 1 µg or 0.5 µg. In this phase, we developed the transfection method with calcium chloride (CaCl2) as an alternative to conventional lipofectamine transfection. The aim was to test whether this more cost-effective method offers comparable transfection efficiency. Three different DNA concentrations were used to investigate the effect on transfection efficiency.
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To enhance transfection efficiency, optimization tests were conducted, in which the quantities of Lipofectamine and DNA were varied. The objective of this iteration was to find the optimal ratio of Lipofectamine 3000 to DNA. To this end, 1 µl and 1.5 µl of Lipofectamine 3000 at a DNA concentration of either 1 µg or 0.5 µg were compared with each other.
To enhance transfection efficiency, optimization tests were conducted, in which the quantities of Lipofectamine and DNA were varied. The objective of this iteration was to find the optimal ratio of Lipofectamine 3000 to DNA. To this end, 1 µl and 1.5 µl of Lipofectamine 3000 at a DNA concentration of either 1 µg or 0.5 µg were compared with each other. In the next step, the tests were carried out with the different DNA concentrations using the CaCl2 transfection method. The transfection efficiencies were compared with those from the Lipofectamine transfection to determine whether the new method represents an improvement.
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The experiment demonstrated that a quantity of 1 µl Lipofectamine 3000 was sufficient for successful transfection, and that increasing the quantity does not result in a notable difference. Additionally, the findings indicated that an amount of 1 µg DNA exhibited a higher efficiency than an amount of 0.5 µg DNA. It can be reasoned that additional factors may have contributed to the previously observed decline in transfection efficiency. One potential explanation is that the cells may have been in an excessively high passage level.
The experiment demonstrated that a quantity of 1 µl Lipofectamine 3000 was sufficient for successful transfection, and that increasing the quantity does not result in a notable difference. Additionally, the findings indicated that an amount of 1 µg DNA exhibited a higher efficiency than an amount of 0.5 µg DNA. It can be reasoned that additional factors may have contributed to the previously observed decline in transfection efficiency. One potential explanation is that the cells may have been in an excessively high passage level. It became clear from the tests that CaCl2 transfection did not deliver better results than Lipofectamine transfection. On the contrary, the efficiency was significantly lower, although the method is less expensive. This led to the realisation that the CaCl2 technique in this form was not a suitable alternative for our specific requirements.
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It can be reasonably deduced that the aforementioned factors may have contributed to the observed decline in transfection efficiency.