MLPA is run on human DNA samples. The main sample requirements are listed below.
Amount: A total quantity of 50-250 ng of DNA in a 5 µl volume is required per MLPA reaction (50-100 ng is optimal). Note that this means that single cells cannot be analysed with MLPA.
Buffer: DNA samples should contain 5-10 mM Tris-HCl buffer with a pH of 8.0-8.5 to prevent depurination during the initial denaturation step. We recommend to dissolve and dilute samples in TE0.1 (10 mM Tris-HCl pH 8.0 + 0.1 mM EDTA). If it is unknown whether sufficient buffering capacity is present, we recommend adding 1 µl of 50 mM Tris-HCl pH 8.5 to 4 μl of sample DNA. Read more about depurination in this article.
Do not dissolve or dilute samples in water, as this lacks sufficient buffering capacity. In addition, do not use PCR buffer, as this contains salts that hamper denaturation.
Tissue source: Suitable DNA sample origin varies between MLPA probemixes but can for example be from blood, fresh healthy/tumour tissue, or FFPE healthy/tumour tissue. Always consult the relevant instructions for use for suitable tissue sources.
Purity: Samples should be free from impurities known to affect MLPA reactions. See this article for more details.
|The instructions for individual probemixes may differ; always read the probemix-specific product description for more details. For more on sample treatment and quality, please read the instructions within the (MS-)MLPA General Protocol.|