General information
The SALSA MLPA Probemix P377 Hematologic Malignancies is a
research use only (RUO) assay, and is intended for screening DNA samples derived from blood or bone marrow for the most common and diagnostically significant copy number changes associated with hematologic malignancies, including acute lymphoblastic leukaemia (ALL), acute myeloid leukaemia (AML), chronic lymphoid leukaemia (CLL), chronic myeloid leukaemia (CML), myelodysplastic syndrome (MDS) and various lymphomas. The probemix can also be used to detect the
JAK2 p.V617F point mutation which is commonly detected in myeloproliferative neoplasm (MPN). This probemix is intended to be used in combination with karyotype analysis. Suggestions on MLPA probemixes that can be used to confirm results or to get a better resolution on genes or chromosomal areas of interest can be found in Table 2 of the product description.
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 P377-B1 Hematologic Malignancies contains 54 MLPA probes with amplification products between 125 and 507 nucleotides (nt). This includes 53 probes for the detection of deletions or duplications in the chromosomal regions 2p (
MYCN,
ALK), 5q (
MIR145,
EBF1 and
MIR146A), 6q, 7p12 (
IKZF1), 7q, 8q24 (
MYC), 9p (
MTAP,
CDKN2A,
CDKN2B,
PAX5), 10q23 (
PTEN), 11q22 (
ATM), 12p13 (
ETV6,
CCND2,
MDM2), 12q, 13q14 (
RB1,
MIR15A,
DLEU2,
DLEU1), 17p13 (
TP53), 17q, Chr 18, Chr 19 and 21q22 (
RUNX1) which are suggested to have a diagnostic or prognostic role in the analysis of samples in hematologic malignancies.
Furthermore, this probemix also contains one probe specific for the
JAK2 p.V617F (c. 1849G>T) point mutation, which will only generate a signal when the mutation is present. In this probemix, 53 out of 54 MLPA probes are used as reference probes, as they are spread over a number of different chromosomal regions and it is expected that the majority of these probes will have a normal copy number in most samples. Complete probe sequences 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 SD068
The SD068 Binning DNA provided with this probemix can be used for binning of all probes including the
JAK2 p.V617F mutation-specific probe (208 nt probe 05672-L17742). SD068 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 SD068 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. 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 SD068 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:
The product description of this Sample DNA can be found on this page under
Online available downloads >
Other product documentation.
Positive samples
Inclusion of a positive sample is usually not required, but can be useful for the analysis of your experiments. MRC Holland has very limited access to positive samples and cannot supply such samples. We recommend using positive samples from your own collection. Alternatively, you can use positive samples from an online biorepository, such as the
Coriell Institute.
See
this support article for commercially available positive samples that have been tested with this product.