MLPA
The MLPA (from English multiplex ligation-dependent probe amplification , dt. Multiplex ligation-dependent amplifying probe ) is a variant of the multiplex PCR for selective propagation and multiplication of DNA sequences such. B. SNPs .
principle
The principle follows the polymerase chain reaction (PCR). In contrast to PCR, the two primers are each separated into two molecules, each half of which has a lower melting temperature than the full-length primer due to its shorter length. The sequences of the primers are chosen so that one of the two primer halves (external or internal) can bind to an area that is identical in all target sequences, while the second primer half can bind to an adjacent area with a different sequence. The primer halves are hybridized with the DNA , followed by ligation . The ligation of two primer halves only takes place in the case of adjacent sequences, which results in two longer primers with a higher melting temperature from the four halves. In a subsequent PCR, due to the annealing temperature used, only DNA that has a ligated primer is amplified. The relative gene dose of related sequences can also be determined using MLPA (as qPCR ) .
Alternatives to MLPA include the DGGE , denaturing HPLC and SSCA ( Single Strand Conformation Analysis ).
Individual evidence
- ↑ Jan P. Schouten, Cathal J. McElgunn, Raymond Waaijer, Danny Zwijnenburg, Filip Diepvens, Gerard Pals1: Relative quantification of 40 nucleic acid sequences by multiplex ligation-dependent probe amplification . In: Nucleic Acids Res . 30, No. 12, 2002, p. E57. doi : 10.1093 / nar / gnf056 . PMID 12060695 . PMC 117299 (free full text).
- ^ SC Yau, M. Bobrow, CG Mathew, SJ Abbs: Accurate diagnosis of carriers of deletions and duplications in Duchenne / Becker muscular dystrophy by fluorescent dosage analysis . In: J. Med. Genet . 33, No. 7, 1996, pp. 550-558. doi : 10.1136 / jmg.33.7.550 . PMID 8818939 . PMC 1050661 (free full text).