Robustness Limitations And Pitfalls In The Use Of Realtime

The use of TaqMan PCR may be particularly difficult if gene expression at a low level is to be quantified. One major pitfall in this context is the accidental determination of genomic DNA when RT-PCR is intended. There are various approaches to meet this problem: It is always useful to select primer combinations that are intron span-ning.[8] If there is no possibility to select intron-spanning primers, RNA samples may be pretreated with DNAse. However, this measure should not be chosen routinely and may also be deleterious if only small amounts of RNA that are partially destroyed as well are present.[9]

One other difficulty is the high degree of technical expertise that is required to achieve as low a variation coefficient and as sensitive a measurement as possible. It could be shown that the degree of technical expertise may alter the gene level that is measured by up to 1000-fold.[9] Because real-time PCR is highly sensitive, the risk of having interference with minor contamination is quite considerable. However, the risk of false-negative results must not be underestimated because post hoc PCR steps are not ''visible'' to the degree that is provided by the

more traditional methods for gene quantification.

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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