Info

SYBR green

Tm analysis

Pathogen detection/differentiation

[18]

SYBR green

Tm analysis

Detection of leukemia specific fusion transcripts

[19]

Minor groove binding probes

Color

SNP detection/genotyping

[20]

that separate the emission spectra. This is particularly critical with increasing degrees of multiplexing in a single tube. The constant generation of new fluorescent dyes, as well as the use of nonfluorescent quenchers (used with 5' nuclease probes, molecular beacons, or minor groove binding probes), has increased the number of different targets that can be discriminated in a reaction. However, considering that there is significant crossover in wavelengths of the emission spectra for most commonly used fluorophores, it is currently practical to distinguish only three or four different colors within a reaction.

When distinguishing between amplified targets on the basis of melting temperatures, salt and magnesium concentrations must be optimized, and Tm values of the probes must be carefully chosen. Because secondary structures and interactions between reaction components cannot always be accurately predicted using computer programs, optimization of reaction conditions and the choice of a proper probe sequence must often be determined empirically. Despite the fact that in theory, dozens of different products can be discriminated in a single reaction by Tm, in reality, for routine clinical assays, a maximum of three or four products is all that can easily be discriminated. This is because of the overlap in Tm ranges; Tm values are relatively consistent, but nevertheless, run to run reaction variability makes it such that a range of at least ±2°C must be allowed between products to ensure that no cross-reactivity is observed.

Getting Started With Dumbbells

Getting Started With Dumbbells

The use of dumbbells gives you a much more comprehensive strengthening effect because the workout engages your stabilizer muscles, in addition to the muscle you may be pin-pointing. Without all of the belts and artificial stabilizers of a machine, you also engage your core muscles, which are your body's natural stabilizers.

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