General information: The SALSA MLPA Probemix P256 FLCN is a research use only (RUO) assay for the detection of deletions or duplications in the
FLCN gene, which is associated with Birt-Hogg-Dube syndrome (BHD). This probemix can also be used to detect the presence of 1285delC and 1285dupC point mutations.
BHD is a rare inherited genodermatosis, characterised by hair follicle hamartomas, kidney tumours, and spontaneous pneumothorax. In a screen of nine BHD families and an additional 53 probands from small BHD families, 27 (44%) were found to have insertion or deletion mutations within a hypermutable C8 tract in exon 11 of the
FLCN gene, located within the Smith-Magenis syndrome region on chromosome 17 (Nickerson et al. 2002,
Cancer Cell, 2002, 2:157-64). Eighteen had a 1-bp insertion and nine had a 1-bp deletion in the C8 tract. A slippage-mediated mechanism during DNA replication is thought to be responsible for these frameshift mutations leading to protein truncation.
More information is available at
https://www.ncbi.nlm.nih.gov/books/NBK1522/.
Probemix content: The SALSA MLPA Probemix P256-B4 FLCN contains 26 MLPA probes with amplification products between 127 and 372 nt. This includes one probe for each of the 14 exons of the FLCN gene and one probe downstream of the FLCN gene. Furthermore, it also contains two probes specific for the 1-bp deletion at 1733C (1285delC, 187 nt probe) and for the 1-bp duplication at 1733C (1285dupC, 194 nt probe) mutations which will only generate a signal when the mutations are present. In addition, nine reference probes are included and detect nine different autosomal chromosomal locations. Complete probe sequences and the identity of the genes detected by the reference probes is available online (
www.mlpa.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, 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.mlpa.com.
SALSA Binning DNA SD032: The SD032 Binning DNA provided with this probemix can be used as Binning DNA sample for binning of two mutation-specific probes (187 nt probe 08598-L08600; 1285delC mutation and 194 nt probe 08598-L08601; 1285dupC mutation). SD032 Binning DNA is a mixture of genomic DNA from healthy individuals and plasmid DNA that contains the target sequences detected by the above mentioned probes. Inclusion of one reaction with 5 μl SD032 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 signals, as for this purpose true mutation positive patient samples or cell lines should be used. It is strongly advised to use DNA sample and reference DNA samples extracted with the same method and derived from the same source of tissue. For further details, please consult the SD032 Binning DNA product description provided.
This product is for research use only (RUO).
sample DNA
Sample DNA developed for this product: