General information
The SALSA MLPA
Probemix P116 SGC is a
research use only (RUO) assay for the detection of deletions or duplications in the
SGCA,
SGCB,
SGCD,
SGCG and
FKRP genes, which are associated with Limb-Girdle Muscular Dystrophy. This probemix can also be used to detect the presence of the
FKRP L276I point mutation.
Limb-Girdle Muscular Dystrophy (LGMD) is characterised by loss of muscle bulk and strength in patients. The distal muscles are affected late in LGMD, if affected at all. LGMD is typically an inherited disorder, though it may be inherited as a dominant, recessive or X-linked genetic defect. The muscle cells of patients with LGMD cannot properly form the proteins needed for normal muscle function. Defects of different proteins are involved in LGMD, each related to a specific type of muscular dystrophy.
Autosomal recessive LGMD is a genetically heterogeneous disorder. Of the many genes that can result in this disorder, the following genes are present in the P116 SGC probemix:
| Gene |
Number of exons |
Number of probes |
Length |
Location |
Distance from p-telomere |
LGMD type |
| SGCA |
10 exons |
11¬ |
9.9 kb |
17q21 |
~46 Mb |
LGMD2D |
| SGCB |
6 exons |
6 |
17.6 kb |
4q12 |
~52 Mb |
LGMD2E |
| SGCD |
9 exons |
9 |
441.0 kb |
5q33 |
~156 Mb |
LGMD2F |
| SGCG |
8 exons |
8 |
144.2 kb |
13q12 |
~23 Mb |
LGMD2C |
| FKRP |
4 exons |
5§ |
12.5 kb |
19q13 |
~52 Mb |
LGMD2I |
¬ This includes a flanking probe located downstream of the
SGCA gene.
§ This includes a probe specific for the
FKRP L276I mutation. This probe will only generate a signal when the mutation is present.
More information is available at
https://www.ncbi.nlm.nih.gov/books/NBK1408/.
This SALSA MLPA probemix is not CE/FDA registered for use in diagnostic procedures. Purchase of this product includes a limited license for research purposes.
Probemix content
The SALSA MLPA Probemix P116-B2 SGC contains 48 MLPA probes with amplification products between 130 and 492 nucleotides (nt). This includes ten probes for the
SGCA gene (one for each exon), one flanking probe downstream of
SGCA, six probes for the
SGCB gene (one for each exon), nine probes for the
SGCD gene (one for each exon), eight probes for the
SGCG gene (one for each exon) and four probes for the
FKRP gene (one for each exon). Furthermore, this probemix contains a probe specific for the
FKRP L276I mutation which will only generate a signal when the mutation is present. In addition, nine reference probes are included that detect autosomal chromosomal locations. Complete probe sequences and the identity of the genes detected by the reference probes are available online (
www.mrcholland.com).
This probemix contains nine quality control fragments generating amplification products between 64 and 105 nt: four DNA Quantity fragments (Q-fragments), two DNA Denaturation fragments (D-fragments), one Benchmark fragment, and one chromosome X and one chromosome Y-specific fragment. More information on how to interpret observations on these control fragments can be found in the MLPA General Protocol and online at
www.mrcholland.com.
SALSA Binning DNA SD030
The SD030 Binning DNA provided with this probemix can be used for binning of all probes including the mutation-specific probe 11373-L13479 (
FKRP L276I mutation). SD030 Binning DNA is a mixture of genomic DNA from healthy individuals and plasmid DNA that contains the target sequence detected by the above mentioned probe. Inclusion of one reaction with 5 μl SD030 Binning DNA in initial MLPA experiments is essential as it can be used to aid in data binning of the peak pattern using Coffalyser.Net software. Furthermore, Binning DNA should be included in the experiment whenever changes have been applied to the set-up of the capillary electrophoresis device (e.g. when capillaries have been renewed). Binning DNA should never be used as a reference sample in the MLPA data analysis, neither should it be used in quantification of mutation signal(s). It is strongly advised that all samples tested are extracted with the same method and derived from the same source of tissue. For further details, please consult the SD030 Binning DNA product description, available online:
www.mrcholland.com.
This product is for research use only (RUO).
Sample DNA
Sample DNA developed for this product: