Journal of Pathogens / 2012 / Article / Tab 5

Research Article

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 ConcentrationReagentFinal ConcentrationAmt/25 uL reactionManufacturer

10xPCR buffer10 mmol−1In beadsGE healthcare
10 mmol−1dNTPs200 umol−1In beadsGE healthcare
50 mmol−1 Mg++1.63 mmol−11.13 uL + beadsInvitrogen
10 umol−1Limiting Primer (4)50 nmol−10.125 uLSigma
100 umol−1Excess Primer (4)1 umol−10.250 uLSigma
10 mmol−1On Probe (7)100 nmol−10.250 uLBiosearch
10 mmol−1Off Probe (4)300 nmol−10.750 uLBiosearch
2.5 U/BeadPure Taq ready-to-go beads1.25 U0.5 beadGE healthcare
106 genome copiesgDNA105 genome copies1 uLATCC