MLPA Flashcards
MLPA
Multiplex Ligation-dependent Probe Amplification
What is MLPA used for?
-uses pairs of probes to detect specific sections of the genome
-used to identify exon-level copy number variation
-a valuable tool for identifying copy number variations (CNVs) associated with various genetic conditions
-Analysing tumour samples
-Studying gene expression (A variation of MLPA called MS-MLPA can be used to analyze DNA methylation)
In simpler terms, MLPA helps researchers see if…
there are extra copies (duplications) or missing copies (deletions) of certain genes or regions in a DNA sample.
By analyzing the number of copies present in a sample, MLPA can aid in diagnosing conditions like:
-Spinal muscular atrophy (SMA)
-Down syndrome
-Cystic fibrosis
-Certain cancers with known CNV associations
MLPA Overview:
-MLPA stands for Multiplex Ligation-dependent Probe Amplification.
-It is a molecular technique used to detect abnormalities in the DNA, such as deletions, duplications, and methylation status.
-It is widely used in genetics and cancer diagnostics.
Key Components of MLPA:
1). Probes: Synthetic oligonucleotides designed to hybridize to specific DNA sequences.
2). Ligation: The process where hybridized probes are ligated (joined) if they are perfectly matched to the target DNA.
3).Amplification: The ligated probes are amplified using PCR (Polymerase Chain Reaction).
MLPA Procedure:
1). DNA Extraction: Isolate DNA from the sample.
2). Hybridization: Probes hybridize to the target DNA sequences.
3). Ligation: Ligase enzyme joins the hybridized probes.
4). Amplification: PCR amplifies the ligated probes.
5). Detection: Amplified products are analyzed using capillary electrophoresis.
MLPA Applications
-Genetic Testing: Detects genetic disorders by identifying deletions or duplications.
-Cancer Research: Identifies gene copy number changes associated with cancer.
-Methylation Analysis: Assesses methylation status of specific genomic regions.
-Carrier Testing: Identifies carriers of genetic mutations.
MLPA Advantages:
Multiplexing: Can analyze multiple targets simultaneously.
Sensitivity: Detects small changes in DNA.
Quantitative: Provides quantitative data on gene copy number.
Limitations of MLPA
Complexity: Requires specific probe design.
Interpretation: Needs careful data analysis and interpretation.
Cost: Can be expensive due to probe synthesis and specialized equipment.
MLPA conclusion
MLPA is a powerful and versatile technique in molecular diagnostics, particularly useful for detecting genetic variations and analysing gene copy numbers.
It combines the benefits of multiplexing and quantitative analysis, making it a valuable tool in both clinical and research settings.
In the MLPA technique, the term “multiplex” refers to
the ability to analyze multiple DNA fragments simultaneously. This feature allows for the efficient detection of changes in gene copy number.
