METHOD FOR MULTIPLEX PCR-ANALYSIS AND SET FOR ITS REALISATION Russian patent published in 2013 - IPC C12Q1/68 G01N33/48 

FIELD: biotechnologies.

SUBSTANCE: method includes using unblocked forward and reverse primers and their homological blocked forward and reverse primers. DNA is separated from a biological or clinical material. The multiplex PCR is carried out with the first 5-10 cycles of amplification in the form of the three-stage PCR and with the following 25-35 cycles of amplification in the form of the double-stage PCR in a buffer for setting of the reaction, containing cations of potassium (I) and cations of magnesium (II), on the basis of the buffer system with pH 7.0-7.5 at 40-85°C, including salt of glutamine acid, tris(hydroxymethyl)aminomethane and orthophosphoric acid. The ratio is selected between quantity of molecules of unblocked primers and quantity of molecules of their homological blocked primers with a non-complementary section from 1-8 nucleotides on the 3'-end, selected from the following row: non-complementary first nucleotide, non-complementary second nucleotide, non-complementary third nucleotide, non-complementary fourth nucleotide, non-complementary fifth nucleotide, non-complementary sixth nucleotide, non-complementary seventh nucleotide, non-complementary eighth nucleotide or their combinations, making from 2:1 to 4:1, when counting the number of the blocked primer nucleotide from the 3'-end.

EFFECT: invention makes it possible to reduce duration of the multiplex PCR without concentration of Taq-polymerase, with prevention of false results related to differences in concentrations of nucleotide sequences-targets.

3 cl, 3 ex