Disadvantages With Mlpa

1. DNA samples should be of equal purity. MLPA reactions are more sensitive to contaminants (PCR inhibitors such as small remnants of phenol) than ordinary PCR reactions. Results on DNA extracted from formaldehyde-treated, paraffin-embedded tissues can be good, but requires a DNA sample from healthy tissue that was treated identically for comparison.

2. It is time-consuming and difficult to develop your own probe mix. Each probe requires the design and preparation of an M13 clone, the purification of single-stranded DNA of that clone, and the digestion of that DNA with expensive enzymes. At this moment, MLPA kits are available only from MRC-Holland. It is possible to design a small number of completely synthetic probes, resulting in amplification products with a length between 100 and 130 nt, and to add these to the MRC-Holland probe mixes. Rules and sequences of the PCR primers are available from [email protected].

3. In contrast to Flourescence in situ hybridization (FISH), MLPA as it is performed today cannot be used to investigate single cells. MLPA analysis of DNA samples from cell mixtures will give the average copy number per cell. It will be difficult to detect deletions of a certain gene if the sample from which the DNA was derived contained less than 60% cancer cells.

4. MLPA is not suitable to detect new mutations, although probes can be made to detect specific known mutations. Mutations or polymorphisms exactly at the ligation site will, in most cases, result in a similar decrease in signal as a deletion of the complete target sequence of that probe. Point mutations and polymorphisms within 4 nucleotides of the ligation site will often affect the signal strength. Point mutations as well as small deletions/insertions within approximately 20 nucleotides of the ligation site may affect the probe signal.

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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