DNA Analyzer Description
MLPA ( Multiplex Ligation-dependent Probe Amplification) method detects multiple copy number changes of genes or loci sites. Nowadays, MLPA is used to check large numbers of hereditary disorders and tumour profiling.Since Dutch Scientist Jan Schouten first invented it, the MLPA method was first published in 2002 'Nucleic Acid Research'. The principle of the MLPA is to apply the specific probe design targeting a region of interest on each sample DNA. MLPA method consists of the following steps:
Denaturation > Hybridization Ligation > PCR MLPA Amplificons Capillary Electrophoresis >Data Analysis
We use sample DNA put into the thermocycler to separate the DNA strands. Then, the targeted DNA strand will be hybridized by adding up the Left and Right probe oligonucleotides. The ligase enzyme will bind the probe oligos up immediately to adjacent target on the sample DNA. Therefore, the hybridized Left and Right probe oligo start catalyzing the creation of a covalent bond in between. Once the probe oligos are sealed, MLPA probes will be exponentially amplified using the single PCR primer pair.
PCR reaction consists of the three steps: denaturation, primer annealing and primer elongation. Those steps of PCR amplification are repeated many times. The fluorescence-labeled primers, which will go through the capillary electrophoresis to pass a detector, are incorporated into the size of the amplification products. The measured fluorescence was visualized as a peak pattern, the so-called electropherogram. The raw data from the capillary electrophoresis analyzer forms the input of the MLPA analysis.
The genetic analyzers of Superyears Gene can be used not only for Sanger sequencing but also for fragment analysis. Fragment analysis First, it obtains the fluorescence-labelled DNA fragments, performs the capillary electrophoresis using a Genetic Analyzer and compares the sample's relative size standard with the designed size standard markers through the analysis software.
The genetic analyzer of Superyears Gene can be used in the capillary electrophoresis during the process of the MLPA experiment, and the data obtained can be used for (compatible) professional analysis software.
The following electropherograms illustrate the targeted samples compared to a set of reference samples and probe ratios of each sample in a ratio chart. Using Superyears Gene's Genetic Analyzer For Mlpa application to analyze Spinal Muscular Atrophy and its results can be visualized. In addition, the Classic series 116 performance of MLPA can also be presented.
Case 1. Spinal Muscular Atrophy (SMA)
MLPA Advantages
MLPA technology combines many features of DNA probe hybridization, PCR technology, and capillary electrophoresis, and develops its ligation-dependent feature.
1.Based upon Sanger Method, Outstanding performance of the golden standard;
2.Support up to 8-color fluorescence, single detection for more than 70 sites;
3.High resolution, High repeatability and Outstanding sequencing quality;
4.Completely open detection platform, compatible with commercial kits in the market;
5.Language interface customizable, free charge of software upgrading;
6.High-cost performance, saving 50%+ of cost in consumables materials and reagents;
7.Certificated by China National Medical Products Administration (NMPA) and CE, ensuring in vitro diagnostic (IVD) applicable;
8.Perfect technical support and after-sale services, one year of free warranty;
MLPA can be applied to:
1. Detect minor rearrangements: BRCA1, BRCA2, MSH2, etc.
2. Detect a wide range of chromosomal rearrangements: Williams syndrome, prill/Angelman syndrome, etc.
3. Detection of copy number changes in subtelomeric regions.
5. Tumor diagnosis: copy number changes in ALL, CLL, Oligodendrogliomas, melanomas, neuroblastomas and other diseases.
6. Quantitative methylation detection: Praderill. angelman syndrome, Fragilex et al.
7. mRNA analysis of apoptosis and inflammatory response.
ALL INFORMATION UPATED ON 12/MAY/2022 BY MARKETING TEAM FROM SUPERYEARS CO.
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