writing a protocol

writing a protocol
AMPLIFICATION
CFX Manager™ Software
Protocol Quick Guide
Run Setup Protocol Tab
The Protocol tab displays a preview of the protocol
loaded in the Run Setup window (Figure 1).
Click Create New to open the Protocol Editor
to create a new protocol.
Click Select Existing to launch the file browser
to load a protocol to use in a run or to edit.
Use the Express Load drop-down menu to directly
load a protocol to use in a run or to edit.
Fig. 1. Protocol tab in the Run Setup window. Load an existing protocol or create a
new protocol for a run.
Click Edit Selected to open the Protocol Editor
to edit the steps of the selected protocol.
Click the Start Run tab to proceed and start a run
with the currently loaded protocol.
Protocol Editor
The Protocol Editor is used to create a new protocol
or edit an existing one (Figure 2).
Graphical
display
1.Select any step in either the graphical or text
display — the step becomes highlighted in blue.
Click on the temperature or dwell time to directly
edit the value.
Text display
2.Click Insert Step to add a temperature step to
the protocol.
Click Delete Step to remove a highlighted
step from the protocol.
3.Click Add Plate Read to Step to designate
when fluorescence data will be acquired during
the protocol. The text on this button changes to
Remove Plate Read if the currently highlighted
step has a plate read. If you do not want to
acquire data at that step, then click Remove
Plate Read.
4.Click on the number of repeats of a GOTO Step
to change the number of cycles in the protocol
(Figure 2, Step 4).
Click on the GOTO Step number to change
the steps included in the GOTO loop.
Protocol
editing
controls
Fig. 2. Protocol Editor. The protocol editing controls are on the
left. Both the graphical and text displays can be edited. Step 1 is
highlighted and 95˚C is selected for editing in the text display.
To add a gradient step:
Graphical display
Gradient range
calculator
1. Click Insert Gradient in the protocol editing
controls (Figure 2).
2. To edit the temperature values for the lowest and
highest temperatures in the gradient, click on the
values in the graphical display, in the text display,
or in the gradient range calculator that appears
to the right of the text display (Figure 3).
3. Within the gradient range calculator, a specific
temperature can be assigned to any row.
Each row is then populated with the appropriate
temperature to satisfy the specified range.
Fig. 3. Insert Gradient step in the Protocol Editor. A gradient step is
highlighted in blue in the graphical and text displays of the protocol. The gradient
range calculator appears when a gradient is added to the protocol.
To add a melt curve:
1. Click Insert Melt Curve in the protocol editing
controls (Figure 2).
2. In the graphical or text display, click on the
temperature values to edit the lowest and
highest temperatures of the melt curve range
(Figure 4, Step 5).
3. Click on the increment value to edit the
temperature interval at which data are acquired.
4. Click on the dwell time to edit the incubation
time for each temperature increment.
Fig. 4. Insert Melt Curve step in the Protocol Editor. A melt curve step is
highlighted in blue in the graphical and text displays of the protocol.
Bio-Rad’s real-time thermal cyclers are licensed real-time thermal cyclers under Applera’s United States Patent Number 6,814,934 B1 for use in
research, human in vitro diagnostics, and all other fields except veterinary diagnostics.
Bio-Rad’s real-time thermal cyclers are covered by one or more of the following U.S. patents or their foreign counterparts owned by Eppendorf AG:
U.S. Patent Numbers 6,767,512 and 7,074,367.
Bio-Rad
Laboratories, Inc.
Web site www.bio-rad.com USA 800 424 6723 Australia 61 2 9914 2800 Austria 01 877 89 01 Belgium 09 385 55 11 Brazil 55 31 3689 6600
Canada 905 364 3435 China 86 20 8732 2339 Czech Republic 420 241 430 532 Denmark 44 52 10 00 Finland 09 804 22 00 France 01 47 95 69 65
Germany 089 31 884 0 Greece 30 210 777 4396 Hong Kong 852 2789 3300 Hungary 36 1 459 6100 India 91 124 4029300 Israel 03 963 6050
Italy 39 02 216091 Japan 03 6361 7000 Korea 82 2 3473 4460 Mexico 52 555 488 7670 The Netherlands 0318 540666 New Zealand 0508 805 500
Norway 23 38 41 30 Poland 48 22 331 99 99 Portugal 351 21 472 7700 Russia 7 495 721 14 04 Singapore 65 6415 3188 South Africa 27 861 246 723
Spain 34 91 590 5200 Sweden 08 555 12700 Switzerland 061 717 95 55 Taiwan 886 2 2578 7189 United Kingdom 020 8328 2000
Life Science
Group
10014290 Rev B
US/EG
10-1333
1110
Sig 1109
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