Академический Документы
Профессиональный Документы
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1. GENERAL INFORMATION
Target genetic element
PCR Assay
Detection Chemistry
TaqMan
Compendium Reference
QT-EVE-ZM-008
2. VALIDATION DATA
Collaborative trial coordinator
JRC-IHCP
Maize flour
Tested GM events
Event Name
NK 603
Unique Identifier
MON-00603-6
Crop Name
Zea mays L.
Method Performance
LOD Relative
0.05%
LOD Absolute
not reported
LOQ Relative
0.1%
LOQ Absolute
not reported
0.10%
0.49%
0.98%
2.0%
4.9%
0.18%
0.85%
1.4%
2.2%
6.0%
RSDr (%)
24%
15%
17%
7.7%
22%
RSDR (%)
37%
34%
25%
26%
31%
Bias %
83.0%
73%
47%
14%
22%
GMO Target
Taxon Target
Mean Slope
not reported
not reported
not reported
not reported
Mean R2
not reported
not reported
Comment
The LOD and LOQ values were provided by the method developer and were not further assessed in the
collaborative trial.
3. REFERENCES
Mazzara M, Paoletti C, Puumalaainen J, Rasulo D, Van Den Eede G. Event-Specific Method for the Quantitation of Maize Line NK603 Using Real-Time PCR - Validation Report and Protocol. EUR 21825 EN. 2005.
JRC32103 (ISBN 92-79-00106-X)
5-ATGAATGACCTCGAGTAAGCTTGTTAA-3
Target element
Insert
Primer Reverse
5-AAGAGATAACAGGATCCACTCAAACACT-3
Target element
3-host genome
Amplicon length
108 bp
Probe
5-FAM-TGGTACCACGCGACACACTTCCACTC-TAMRA-3
Target element
Taxon-target(s)
Primer Forward
5-CCAGCCTCATGGCCAAAG-3
Target element
adh1
Primer Reverse
5-CCTTCTTGGCGGCTTATCTG-3
Target element
adh1
Amplicon length
70 bp
Probe
5-FAM-CTTAGGGGCAGACTCCCGTGTTCCCT-TAMRA-3
Target element
Taxon-target(s)
Final Concentration
Reagent
Final Concentration
1x
1x
Primer Fw
0,15 mol/L
Primer Fw
0,15 mol/L
Primer Rev
0,15 mol/L
Primer Rev
0,15 mol/L
Probe
0,05 mol/L
Probe
0,05 mol/L
Nuclease-free water
Nuclease-free water
Template DNA
maximum 300
Template DNA
maximum 300
Final Volume
50 L
Final Volume
50 L
6. AMPLIFICATION CONDITIONS
GM-target(s) and taxon-target(s)
Stage
Temperature
Time
No Cycles
Decontamination (UNG)
50C
120
Activation/Initial Denaturation
95C
600
Denaturation
95C
15
60C
60
45