General information
The SALSA MLPA
Probemix P175 Tumour Gain is a
research use only (RUO) assay for the detection of copy number aberrations in 24 genes, which are frequently gained or amplified in various tumour types. This probemix can also be used to detect the presence of the
BRAF p.V600E (c.1799T>A) point mutation.
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 P175-B1 Tumour Gain contains 62 MLPA probes with amplification products between 115 and 504 nucleotides (nt). This includes two probes for each of the following genes:
ABL1,
ALK,
AR, AURKA/B,
BRAF,
CCND1/2,
CDK4,
DHFR,
EGFR,
ERBB2,
FGFR1,
KDR,
KIT,
MDM2/4,
MET,
MYC,
MYCN,
PDGFRA,
RET,
SMO and
TOP2A. Furthermore, this probemix also contains one probe specific for the
BRAF p.V600E (c.1799T>A) point mutation which will only generate a signal when the mutation is present. In addition, 13 reference probes are included that detect autosomal chromosomal locations which are relatively stable in most tumour types. However, it should be noted that tumour karyotypes can harbour multiple numerical and structural aberrations, which can complicate interpretation of these reference probes. Complete probe sequences and the identity of the genes detected by the reference probes are available in Table 3 of the Product Description and 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 SD029
The SD029 Binning DNA provided with this probemix can be used for binning of all probes including the one mutation-specific probe (
BRAF probe 08780-SP0039-L08904 for the p.V600E (c.1799T>A) point mutation). SD029 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 SD029 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 the mutation signal. 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 SD029 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: