Chapter 3 Prepare the Relative Standard Curve Reactions
Prepare the Reaction Plate
61
Applied Biosystems 7500/7500 Fast Real-Time PCR System Getting Started Guide for Relative Standard Curve
and Comparative C
T
Experiments
Notes
Required
Materials
• Microcentrifuge tubes
• Pipettors
• Pipette tips
• c-myc reaction mix (from page 59)
• GAPDH reaction mix (from page 59)
•Water
• c-myc standards (from page 55)
• GAPDH standards (from page 56)
• Samples (from page 54)
•MicroAmp
®
Optical 96-Well Reaction Plate
•MicroAmp
™
Optical Adhesive Film
• Centrifuge
Prepare the
Reaction Plate
1. For each target, prepare the negative control reactions:
a. To an appropriately sized tube, add the volumes of reaction mix and water
listed below.
b. Mix the reaction by gently pipetting up and down, then cap the tube.
c. Centrifuge the tube briefly to remove air bubbles.
d. Add 50 µL of the negative control reaction to the appropriate wells in the
reaction plate.
Tube Reaction Mix
Reaction Mix
Volume (µL)
Water Volume
(µL)
1 c-myc reaction mix 59.4 6.6
2 GAPDH reaction mix 59.4 6.6
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