- Which Technique Is More Accurate Why Fluorometric Photometry Is More Accurate Than Mini Prep Method Due To Wider Conc 1 (80.58 KiB) Viewed 43 times
- Which Technique Is More Accurate Why Fluorometric Photometry Is More Accurate Than Mini Prep Method Due To Wider Conc 2 (457.52 KiB) Viewed 43 times
Fluorometer 1. A fluorometer or fluorimeter is a device used to measure parameters of fluorescence: its intensity and wavelength distribution of emission spectrum after excitation by a certain spectrum of light 2. These parameters are used to identify the presence and the amount of specific molecules in a medium. Modern fluorometers are capable of detecting fluorescent molecule concentrations as low as 1 part per trillion. Miniprep Minipreparation of plasmid DNA is rapid, small-scale isolation of plasmid DNA from bacteria. It is based on the alkaline lysis method. The extracted plasmid DNA resulting from performing a miniprep is itself often called a "mini-prep". Minipreps are used in the process of molecular cloning to analyze bacterial clones. A typical plasmid DNA yield of a miniprep is 50 to 100 ug depending on the cell strain. Miniprep of a large number of plasmids can also be done conveniently on filter paper by lysing the cell and eluting the plasmid on to filter paper. Answer-Fluorometric Photometry is more accurate than mini prep Wide concentration range: Fluorimetry generally can detect more than three to six log orders of concentration without sample dilution or modification of the sample. Accurate results: The sensitivity and specificity of fluorescence measurement lead to potentially more precise and accurate readings. Answer - Mini prep contamination with unknown material If your plasmid is 4 kbps and your unexpected band is 1.5 to 2 kbps, there are two possible scenarios: it could be that your plasmid is probably broken during the miniprep process, or you might have a piece of the broken genomic bacterial DNA in your miniprep product. If you are sure that the pattern that you get from gel electrophoresis of the restricted fragments of this unexpected band does not match with the plasmid sequence in any way, then you have DNA contamination, very likely from the host bacteria. -Fluorometric Photometry discrepancies 1. If the sample is turbid, this will result in an error as the light will be scattered and part of it will never reach the detector and thus will appear as absorbed. 2. If the molecule is capable of association/dissociation and the absorbances of the two forms differ, the Lambert-Beer law will no longer be valid as the degree of dissociation will depend on the concentration. 3. It is crucial for the cuvette to be clean and free of scratches. The sides of the cuvette in the light path should never be touched by hand. Answer - Integrity of plasmid could be well understood by the Gel Miniprep as Minipreparation of plasmid DNA is rapid, small-scale isolation of plasmid DNA from bacteria. It is based on the alkaline lysis method. The extracted plasmid DNA resulting from performing a miniprep is itself often called a "mini-prep". Minipreps are used in the process of molecular cloning to analyze bacterial clones. while A photometer is a tool used to measure light, and photometry is how light is measured. ... Specifically, photometers can measure ultraviolet to infrared on the electromagnetic spectrum, which includes visible light. Photometry is used in chemistry to help identify what materials are in substances.