Multiplex PCR Master Mix (2x) is developed for efficient multiplex PCR for various advanced applications. The use of the kit eliminates the need for time-consuming optimization, making the development of multiplex PCR assays easier and faster. Use of Multiplex PCR Master Mix (2x) allows to save time and reduce contamination risk due to fewer pipetting steps during PCR setup. The master mix contains Perpetual Taq DNA Polymerase, MgCl2, specially formulated reaction buffer, dNTPs and additives. Use of the “hot start” enzyme prevents extension of misprimed products and primer-dimers during reaction setup leading to higher specificity and sensitivity of PCR reactions. Multiplex PCR Master Mix (2x) enables convenient room temperature reaction setup. The master mix can be used with plain primers or fluorescently labeled primers.