Techne PrimeQ User Manual Page 135
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- Table of contents
- TROUBLESHOOTING
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3.15.1 Assay requirements
•
Need at least one reporter dye.
•
Need a stage in the program with a ramp read (with at least two readings).
3.15.2 Setup
•
Program a ramp read into the PCR program as described in section 3.4.3.6.
•
In the Analysis Selection box in the Experiment Editor, click on the name of the ramp stage
and then press
Edit
.
•
This will lead through to the Analysis Wizard Selection box. Select
Dissociation Curve
in
the drop-down menu and select dye usage.
•
Click
Next
and the Dissociation Curve Wizard will launch.
The wizard will lead the user through the setup:
•
Background correction:
Corrects for effects of temperature on fluorescence and the
background before and after all the DNA has melted.
•
Digital filter:
Smoothes the dissociation curve raw data (but does not affect the calculated
results).
•
Peak detector:
Finds specified number of peaks and records the dissociation temperature
(Tm).
•
Peak area:
Calculates the area under the peaks detected.
3.15.2.1 Background correction
The first window of the wizard allows setting of background correction parameters.
Raw data
for reporter
Background
correction and
optional curve
smoothing filter
Dissociation
curve
Dissociation
peaks
Report
Tm Peak areasRaw data
for reporter
Background
correction and
optional curve
smoothing filter
Dissociation
curve
Dissociation
peaks
Report
Tm Peak areas
- OPERATOR’S MANUAL 1
- How to use this guide 3
- Contents 7
- 1 Introduction 13
- 1.1 PrimeQ 15
- 1.2 Quansoft 15
- 1.2.3 Program Editor 16
- 1.2.4 PrimeQ reports 16
- 1.2.5 Data Analysis 16
- 1.3 Unpacking 17
- 1.4 Warning! 17
- 1.5 Uses of PrimeQ 17
- 1.6 Plates 17
- 1.7 Thermal seal 18
- 1.8 Contacting us 18
- 1.9 Installing the hardware 18
- 1.9.3 Operator safety 20
- 1.9.4 Installation 20
- 1.10 Before switching on 23
- 1.11 Technical Specification 24
- 1.12 Working conditions 25
- 1.13 Guarantee 25
- WARRANTY 26
- IF BROKEN 26
- 1.14.4 Software upgrades 29
- 1.15.1 Function keys 30
- 1.15.2 Power up screen 30
- 1.15.3 Beeper 30
- 1.15.4 Before you start 31
- 1.15.5 Start-up procedure 31
- 1.16.8 Points to remember 35
- 1.18 LED power settings 37
- 1.19 Inserting a plate 37
- 1.20 After use 38
- 2 Getting started 39
- 2.1 Introduction to PrimeQ 41
- 2.2.1 PCR 42
- 2.3.1 Intercalating dyes 44
- 2.3.3 Molecular Beacons 45
- 2.4 System overview 46
- 2.5 Optics module 46
- 2.6 Thermal cycling module 47
- 2.7 Scanning module 48
- 3 Quansoft 49
- 3.1 Introducing Quansoft 51
- 3.2 Software overview 52
- 3.2.1 Home page 53
- 3.2.2 The Editors 53
- 3.2.2.3 Program Editor 54
- 3.2.2.4 Results Editor 54
- 3.3 Using Quansoft 55
- 3.3.2 Main screen 56
- 3.3.3 Navigation bar 56
- 3.3.4 Title bar functions 57
- 3.3.5 Menu bar functions 58
- 3.3.6 Accessing the editors 59
- 3.4.1 An Overview 60
- 3.4.3 Setting up a program 62
- Comment [MO1]: 65
- Discussed between 65
- Matt and Christian 65
- 3.5 Running an experiment 86
- 3.5.2 Monitoring the run 89
- 3.6 LED intensity settings 92
- 3.7 Results Editor 93
- 3.7.3 Editing a graph 96
- 3.7.5 Log/Audit trail 99
- 3.7.6 Report 99
- 3.8.1 Exporting 101
- 3.8.2 Printing 102
- Data analysis 103
- 3.9 Introduction 105
- 3.9.3 Fit points 106
- -1/slope 107
- 3.11 Analysis method: None 111
- 3.11.2 PrimeQ Report 112
- 3.13.1 Assay requirements 114
- 3.13.2 Setup 114
- 3.13.3 Viewing the results 116
- 3.13.4 PrimeQ Report 118
- 3.14.1 Assay requirements 119
- 3.14.2 Setup 119
- Crossing line 121
- Noise threshold 121
- 3.14.3 Viewing the results 123
- 3.14.4 PrimeQ Report 124
- 3.14.5 Using Cq values 124
- R² = 0.996 E = 1.977 126
- Dissociation curve 134
- 3.15.1 Assay requirements 135
- 3.15.2 Setup 135
- 3.15.3 Viewing the results 141
- 3.15.4 PrimeQ Report 142
- 3.16.1 Assay requirements 144
- 3.16.2 Setup 144
- 3.16.3 Viewing the results 146
- The plus-minus graph 147
- Results table 147
- 3.16.4 PrimeQ Report 148
- 3.17.1 Assay requirements 150
- 3.17.2 Setup 150
- 3.17.3 Viewing the results 152
- Reporter Selection 153
- 3.17.4 PrimeQ Report 155
- 3.18.1 Assay requirements 157
- 3.18.2 Setup 157
- 3.18.3 Viewing the results 158
- Multi-read graph 159
- 3.18.4 PrimeQ Report 160
- 3.19.1 Multiplex setup 162
- 4 Technical information 165
- 4.1.1 Cleaning PrimeQ 167
- 4.1.2 Fuses 167
- 4.1.3 Insulation Testing 168
- 4.1.4 Mantenimiento 168
- 4.2.1 LED 169
- 4.2.2 Filters 169
- 4.2.3 Replacing the fuses 169
- 4.5 Accessories 170
- 4.6 Replacement parts 171
- 4.8 Packaging 174
- 4.9 Trademarks 174
- 5 Troubleshooting 175
- 5.1 Troubleshooting 177
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