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Fig. 2 RFLP analysis of c-kit. After amplification of the mutation-containing region of the c-kit cDNA, PCR products were digested with Hinfi and analyzed by electrophoresis on a polyacrylamide gel. Compared to WT c-kit (1), the c-kit D816V mutation (2) generates a second restriction site for the endonuclease Hinfl within the amplified region. Predicted sizes for the WT sequence are 150 and 56 bp, and are 150, 42, and 14 bp for the mutated sequence.

approach combining peptide nucleic acid-mediated polymerase chain reaction (PCR) clamping and detection of c-kit mutations with hybridization probes has been described.1-25-1 The major advantage of this method is that the mutant allele is amplified selectively and can be detected in case of a more than 1000-fold excess of WT over mutated c-kit. In addition, sequence variations other than the D816V c-kit mutation can be detected within the region covered by the probes. However, all these techniques are not sufficient to identify the cell type (MCs vs. other cells in tissues or blood) in which the mutation is present. To address this subject, isolation of the cell type(s) of interest is required. One possible approach is to collect immunohistochemically defined subpopulations of cells from stained tissue sections by microdissection.1-24-1 An alternative strategy is sorting of cell subpopulation from BM or peripheral blood (PB) by flow cytometry.[26] Using these techniques, even single cells can be isolated and analyzed for c-kit D816V.[24,26] However, these techniques are not used in the routine workup of patients with (suspected) SM. Rather, RFLP analysis of D816V is considered to be appropriate and sufficient for routine diagnostic purposes.

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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