Bio-Rad Precision Melt Analysis™ Software - Manual (Page 16)

Precision Melt Analysis Software Instruction Manual
11
Preparing the Melt Calibration Plate
Bio-Rad Laboratories provides the following materials in the Melt Calibration kit (184-5020) to
create a melt calibration plate:
• 10016289. Melt Calibration DNA Standard
• 10016273. Melt Calibration primers
• 1725110. Precision Melt supermix
Additional Materials Required
In addition to the components provided in the melt calibration kit, the following materials are
required to prepare the melt calibration plate:
• PCR-grade tubes and nuclease-free water
• MSB-1001. Microseal
®
‘B’ adhesive seals, optically clear
To calibrate a CFX384 system:
• HSP-3805. Hard-Shell
®
thin-wall 384-well skirted PCR plates with clear shell and white
wells for use with a CFX384 system
To calibrate a CFX96 or CFX Connect system:
Choose white wells or natural wells depending on the plate type you plan to use in your
experiments
• HSP-9601. Hard-Shell thin-wall 96-well skirted PCR plates with white shell and clear
wells
• HSP-9655. Hard-Shell thin-wall 96-well skirted PCR plates with white shell and white
wells
WARNING! Precision melt supermix is stable for 6 months when stored in a
constant temperature freezer at –20°C, protected from light. For convenience, it
may be stored at 2–8°C for up to 3 months. Repeated freezing and thawing of the
supermix is not recommended.
Preparing the melt calibration plate
1. Add the required volumes of each component to an appropriately sized tube (Table 5).
2. Cap the tube and mix the reaction components gently by vortexing.
3. Briefly centrifuge the tube to remove air bubbles and collect contents at the bottom of
the tube.
4. Add the appropriate volume of the mixture into each well of a reaction plate
• For a 96-well system, add 20 μl to each well of a 96-well plate.
• For a 384-well system, add 10 μl to each well of the 384-well plate
Table 5. Reaction set up for melt calibration plate.
Component
Volumes (μl) for
96-well System Volumes (μl) for
384-well System
Precision Melt Supermix 1,200 2,250
Melt Calibration DNA Standard 120 450
Melt Calibration Primers 14.4 27
PCR-Grade Water 1065.6 1,773
Total 2,400 4,500