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SALSA® digitalMLPA

SALSA® digitalMLPA is similar to conventional MLPA, the gold standard for CNV detection of multiple DNA sequences, but with the ability to examine many more targets. digitalMLPA takes the guesswork out of NGS CNV calls by combining the MLPA technology you trust for CNV detection with the large number of targets you have come to expect with NGS. With digitalMLPA, you can examine more targets than with conventional MLPA, with decreased costs and turnaround times.


SALSA® digitalMLPA – where MLPA meets NGS

The fastest way to broad copy number certainty
✓ Reliable: proven copy number detection technology+
✓ Robust: only 20 ng of sample DNA needed; uniform coverage
✓ Quality control: control probes included in each reaction
✓ Simple: easy hands-on steps and no library quantification needed
✓ Cost-effective: up to 1000 probes in one reaction
✓ Sample certainty: SNV probes present to distinguish samples
✓ Easy analysis: user-friendly analysis with free Coffalyser digitalMLPA software

SALSA® digital Multiplex Ligation-dependent Probe Amplification (digitalMLPA) is a
multiplex PCR followed by Illumina sequencing-based amplicon quantification, for the
detection of copy number variations (CNV) and specific point mutations. digitalMLPA
amplifies ligated probes with a universal primer pair enabling unbiased amplification.
With digitalMLPA, up to 1000 unique sequences can be detected and quantified in a
single reaction. 

digitalMLPA samples can be combined with other NGS sequencing libraries in a single run to give simultaneous results for reliable CNV quantification and NGS sequence analysis.

This saves time and money, ensuring sample result turnaround times are met.

digitalMLPA data analysis is count-based, uses free software (Coffalyser digitalMLPA),
and can be done on any Windows 10-based personal computer. Coffalyser digitalMLPA returns two reports for easy reaction quality determination and result interpretation. Best of all, no bioinformatician is needed for result interpretation.

digitalMLPA workflow

FIRST digitalMLPA ASSAY ON THE MARKET in 2020: SALSA digitalMLPA Probemix D001 Hereditary Cancer Panel 1


To perform a digitalMLPA reaction, three items are required: (1) a SALSA digitalMLPA probemix, (2) a SALSA digitalMLPA reagent kit (DRK), and (3) a SALSA barcode solution plate (two different plates of 96 barcode solutions are available). Currently digitalMLPA barcode solution plates and reagent kits are only compatible with Illumina instruments.


digitalMLPA, like conventional MLPA, is based on the sample DNA-dependent generation of ligated probe products, followed by PCR amplification of ligated probes by a single PCR primer pair (Schouten JP et al. (2002) Nucleic Acids Res. 30:e57, Figure 1). In digitalMLPA, up to 1000 different probes can be included in a probemix and used in a single reaction.

Data analysis by Coffalyser digitalMLPA first determines the absolute read number for each probe by assigning each read to a sample using the unique barcode and to a specific probe by the unique probe sequence. Next, the absolute read counts of a sample are converted into relative values (intra ratios) by normalising target probe read counts against the read counts of every reference probe in the sample (intra-normalisation). This is done for every probe and every sample. In the next step, called internormalisation, Coffalyser digitalMLPA compares the relative probe values of each sample to those of the reference samples, resulting in inter ratios. (source: SALSA® digitalMLPA General Protocol)

digitalMLPA products are for research use only (RUO).

Videók, ismertetők

SALSA® digitalMLPA Confidence in Copy Number Determination (pdf)

SALSA® digitalMLPA General Protocol (pdf) - for use with Illumina sequencing platforms

digitalMLPA Protocol Supplements (pdf) - Running digitalMLPA reactions on NextSeq 500/550 instruments and Combining digitalMLPA reactions and standard paired-end sequencing reactions.

For detailed informations and latest versions of protocols and product descriptions, please visit


What is digitalMLPA?

digitalMLPA is a technique for the copy number quantification of up to 1000 target sequences. The technique combines the conventional MLPA technology with next-generation sequencing (NGS). This video briefly describes what digitalMLPA is and what steps are involved.


SALSA digitalMLPA Probemix D001 Hereditary Cancer Panel 1

D001 Hereditary Cancer Panel 1 genes covered

The digitalMLPA probemix D001 detects CNVs in 28 clinically relevant genes, and detects six specific common (point) mutations. Mutations in these genes are associated with hereditary predisposition to one or more of the following cancer types: breast, ovarian, colorectal, gastric, prostate, pancreatic, endometrial, and melanoma. Read more about the targeted genes on the product page.

Save time and money on NGS-negative samples

D001 is the perfect time-saving complement to NGS sequencing for high-level CNV calling certainty. digitalMLPA is highly multiplexable, where digitalMLPA reactions and NGS libraries can be run on the same chip. This enables simultaneous CNV quantification and NGS sequence analysis, avoiding extra copy number tests run on NGS-negative samples.

Tartozékok, kiszerelések

Coffalyser digitalMLPA

Coffalyser digitalMLPA is free software developed by MRC Holland for the analysis of digitalMLPA data. Coffalyser digitalMLPA automatically recognises digitalMLPA sequence reads from FASTQ files and directly uses them for analysis. Coffalyser digitalMLPA then outputs two clear reports for every sample of your experiment indicating both the quality of the run and the aberrations found.


Coffalyser digitalMLPA is for research use only (RUO).



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