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When using spectrophotometric analysis to determine the concentration of DNA or RNA, the Beer–Lambert law is used to determine unknown concentrations without the need for standard curves. In essence, the Beer Lambert Law makes it possible to relate the amount of light absorbed to the concentration of the absorbing molecule.
The Qubit fluorometer method is to use fluorescent dyes to determine the concentration of either nucleic acids or proteins in a sample. Specialized fluorescent dyes bind specifically to the substances of interest. A spectrophotometer is used in this method to measure the natural absorbance of light at 260 nm (for DNA and RNA) or 280 nm (for ...
The C 0 t value is the product of C 0 (the initial concentration of DNA), t (time in seconds), and a constant that depends on the concentration of cations in the buffer. Repetitive DNA will renature at low C 0 t values, while complex and unique DNA sequences will renature at high C 0 t values. The fast renaturation of the repetitive DNA is ...
Damage to DNA – Exposure of DNA to UV radiation in standard preparative agarose gel electrophoresis procedure for as little as 45 seconds can damage the DNA, and this can significantly reduce the transformation efficiency. [19] Adding cytidine or guanosine to the electrophoresis buffer at 1 mM concentration however may protect the DNA from ...
Nucleotide bonds showing AT and GC pairs. Arrows point to the hydrogen bonds.. In molecular biology and genetics, GC-content (or guanine-cytosine content) is the percentage of nitrogenous bases in a DNA or RNA molecule that are either guanine (G) or cytosine (C). [1]
Several formulas are used to calculate T m values. [10] [11] Some formulas are more accurate in predicting melting temperatures of DNA duplexes. [12] For DNA oligonucleotides, i.e. short sequences of DNA, the thermodynamics of hybridization can be accurately described as a two-state process.
The DNA is digested by a particular restriction enzyme, resulting in DNA pieces of varying molecular masses. One of the advantages of this method is that more marker can readily be created simply by digesting more of the known DNA. [3] On the other hand, the size of the DNA pieces are based on the sites where the restriction enzyme cuts.
The RFU measurements are used, for DNA profiling, in a real-time polymerase chain reaction (PCR). Two common methods for detection of products in real-time PCR are: (1) non-specific fluorescent dyes that intercalate with any double-stranded DNA, and (2) sequence-specific DNA probes consisting of oligonucleotides that are labeled with a fluorescent reporter which permits detection only after ...