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Table 5 PRI-lock assay validation. Analysis of target DNAs mixed in different ratios.

From: Quantitative multiplex detection of plant pathogens using a novel ligation probe-based system coupled with universal, high-throughput real-time PCR on OpenArrays™

Template (Log10 target

(copy number/μl ligation reaction))

Ratio

Observed CT values (SD) in PRI-lock specific PCR

Observed pathogen template (SD)

(Log10 target (copy number/μl ligation reaction))

P. sojae

E. carotovora carotovora

 

Phytophthora spp.

E. carotovora carotovora

Phytophthora spp.

E. carotovora carotovora

8

5

1000: 1

9.4 (0.30)

19.3 (0.23)

7.9 (0.08)

4.9 (0.07)

8

4

10000: 1

9.3 (0.06)

22.7 (0.34)

8.0 (0.02)

3.9 (0.10)

5

8

1: 1000

20.6 (0.20)

8.8 (0.14)

4.8 (0.06)

7.9 (0.04)

4

8

1: 10000

24.2 (0.80)

8.9 (0.07)

3.8 (0.22)

7.8 (0.02)

  1. Target numbers were calculated using the CT values and the corresponding calibration curves. Data represent 2 ligation replicates (n = 2), each with 4 qPCR replicates (n = 4).