Standardisation and evaluation of a quantitative multiplex real-time PCR assay for the rapid identification of Streptococcus pneumoniae

Table 2 Quantitative multiplex real-time PCR (qmPCR) set-up

Component	Volume/reaction	Final concentration
2X Rotor-Gene Multiplex PCR Master Mix	12.5 µl	1X
20X primer-probe mix *5^a	1.25 µl*5	0.5 µM forward primer*5
		0.5 µM reverse primer *5
		0.2 µM probe *5
Template DNA	5 µl	<100 ng/reaction
DNase/RNase free water	1.25 µl
Total reaction volume	25 µl

^a20X primer-probe mix for all 5 primer-probe sets was prepared and added individually in a single-reaction

ISSN: 2200-6133