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.

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

digitalMLPA ASSAY COMPONENTS

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 ASSAY PRINCIPLE

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).