Thermo Fisher Scientific TaqMan® Pathogen Owner's manual

Brand
Thermo Fisher Scientific
Model
TaqMan® Pathogen
Type
Owner's manual
STEP
5 µL Negative Control
ACTION
TaqMan
®
Pathogen
Detection Kit
Reaction Setup
A. Make the Pathogen PCR Cocktail ( )
as shown in this table.
B. Dispense 45 µL of the Pathogen PCR Cocktail
into all of the wells except the NAC wells.
This example shows 30 UNKN wells.
• Add
into each of the NTC wells.
• Cap the 1st column of the plate
with Optical caps.
A
B
C
D
E
F
G
H
123456789101112
• Add 5 µL of your sample DNA
into each of the UNKN ( ) wells.
• Cap all of the wells except those in column 12 .
P/N 4307908 Rev. A
Number of
Unknowns
TaqMan
Pathogen
Mix (µL)
AmpliTaq Gold®
DNA Polymerase
(µL) Total (µL)
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
880
1144
1364
1628
1848
2112
2332
2596
2816
3080
3300
3564
3784
4048
4268
20
26
31
37
42
48
53
59
64
70
75
81
86
92
97
900
1170
1395
1665
1890
2160
2385
2655
2880
3150
3375
3645
3870
4140
4365
A
B
C
D
E
F
G
H
123456789101112
NTC
44 µL TaqMan Pathogen Mix
5 µL Negative Control
• Add
into the six NAC wells.
A
B
C
D
E
F
G
H
123456789101112
NAC
A
B
C
D
E
F
G
H
123456789101112
No Template Control
No Amplification
Control
Pos
Ctr
5 µL Positive Control
• Add
into the NAC wells.
• Add
into each of the two positive control wells.
• Cap the wells in column 12.
A
B
C
D
E
F
G
H
123456789101112
Pos Ctr
TaqMan
®
Pathogen
Detection Kit
Procedure Flowchart
See Product Insert for details.
See the PrepMan Protocol (P/N 4305840) for details.
See reverse side of this card for details.
Open the Plate-Read stationery file
Set up the plate configuration
Label the positive control wells
Name the sample wells
Add a comment in the plate setup window
Save the file. This is the plate-read data file
P/N 4307908 Rev. A
Optional
Optional
Instrument Setup
Reaction Setup
Pre-Read
Post-Read
Analysis
PCR Thermal Cycling
PrepMan™ Sample
Preparation Reagent
DNA Purification
Pre-Enrichment
Sample
9 min.
94˚
20 sec.
95˚
Stage 1 Stage 2
1 min.
60˚
Repeat 40
30 sec.
72˚
In analysis plate window, click Pre-Read
Save the pre-read file. This is the plate-read data file
Check the Raw Spectra view for the pre-read
Open the Thermal Cycling stationery file
Click show analysis button
Click the Run button in the Analysis plate window
Save the data. This is the real-time data file
Open the file containing the pre-read data
Click Show Analysis
Click the Post-PCR read on the Analysis plate window
Save the data. This is the plate-read file
Open the plate-read file
Choose Analyze from the Analysis menu
Print the Event log dialog box
Select Experiment Report from the Window menu
Print the Experiment Report
16–24 hrs
0.5–2 hrs
15–20 min
15–60 min
2 min
2 min
2 min
2.5 hrs
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Thermo Fisher Scientific TaqMan® Pathogen Owner's manual

Brand
Thermo Fisher Scientific
Model
TaqMan® Pathogen
Type
Owner's manual
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