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NetPrimer is a gratis web-based tool used for analysing primers used in PCR to amplify a DNA sequence. [2] The software predicts the melting temperature of the primers using the nearest neighbor thermodynamic algorithm. The accurate prediction of the melting temperature (Tm) is one of the most important factors that governs the success of a PCR ...
Only a narrow concentration of BSA is used (2-10 ug/mL) in order to create an accurate standard curve. [23] Using a broad range of protein concentration will make it harder to determine the concentration of the unknown protein. This standard curve is then used to determine the concentration of the unknown protein.
There are two common methods in which to construct a DNA molecular-weight size marker. [3] One such method employs the technique of partial ligation. [3] DNA ligation is the process by which linear DNA pieces are connected to each other via covalent bonds; more specifically, these bonds are phosphodiester bonds. [4]
When using a path length that is shorter than 10mm, the resultant OD will be reduced by a factor of 10/path length. Using the example above with a 3 mm path length, the OD for the 100 μg/mL sample would be reduced to 0.6. To normalize the concentration to a 10mm equivalent, the following is done: 0.6 OD X (10/3) * 50 μg/mL=100 μg/mL
A primer dimer (PD) is a potential by-product in the polymerase chain reaction (PCR), a common biotechnological method. As its name implies, a PD consists of two primer molecules that have attached to each other because of strings of complementary bases in the primers.
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The free NCBI tool Primer-BLAST integrates primer design and BLAST search into one application, [6] as do commercial software products such as ePrime and Beacon Designer. Computer simulations of theoretical PCR results ( Electronic PCR ) may be performed to assist in primer design by giving melting and annealing temperatures, etc. [ 7 ]
Plasmid miniprep. 0.8% agarose gel ethidium bromide-stained.. A plasmid preparation is a method of DNA extraction and purification for plasmid DNA.It is an important step in many molecular biology experiments and is essential for the successful use of plasmids in research and biotechnology.