Design and Construction of a Single-Tube, LATE-PCR, Multiplex Endpoint Assay with Lights-On/Lights-Off Probes for the Detection of Pathogens Associated with Sepsis

Table 5

LATE-PCR 16S sepsis multiplex reagents.

Starting Concentration	Reagent	Final Concentration	Amt/25 uL reaction	Manufacturer
10x	PCR buffer	10 mmol^−1	In beads	GE healthcare
10 mmol^−1	dNTPs	200 umol^−1	In beads	GE healthcare
50 mmol^−1	Mg^++	1.63 mmol^−1	1.13 uL + beads	Invitrogen
10 umol^−1	Limiting Primer (4)	50 nmol^−1	0.125 uL	Sigma
100 umol^−1	Excess Primer (4)	1 umol^−1	0.250 uL	Sigma
10 mmol^−1	On Probe (7)	100 nmol^−1	0.250 uL	Biosearch
10 mmol^−1	Off Probe (4)	300 nmol^−1	0.750 uL	Biosearch
2.5 U/Bead	Pure Taq ready-to-go beads	1.25 U	0.5 bead	GE healthcare
10^6 genome copies	gDNA	10^5 genome copies	1 uL	ATCC