Applied Biosystems 7500 Fast Getting Started

Chapter 3 Prepare the Reactions
Prepare the Reaction Plate
Applied Biosystems 7500/7500 Fast Real-Time PCR System Getting Started Guide for Standard Curve
Experiments
52
Notes
Required
Materials
• Microcentrifuge tubes
• Pipettors
• Pipette tips
• RNase P reaction mix (from page 49)
•Water
• Standards (from page 47)
• Samples (from page 45)
•MicroAmp
®
Optical 96-Well Reaction Plate
•MicroAmp
™
Optical Adhesive Film
• Centrifuge
Prepare the
Reaction Plate
1. Prepare the negative control reactions for the target:
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 25 µL of the negative control reaction to the appropriate wells in the
reaction plate.
2. For each replicate group, prepare the standard reactions:
a. To appropriately sized tubes, add the volumes of reaction mix and standard
listed below.
b. Mix the reactions by gently pipetting up and down, then cap the tubes.
c. Centrifuge the tubes briefly to remove air bubbles.
d. Add 25 µL of the standard reaction to the appropriate wells in the reaction plate.
Tube Reaction Mix Reaction Mix Volume (µL) Water Volume (µL)
1 RNase P reaction mix 74.25 8.25
Tube Standard Reaction Reaction Mix
Reaction Mix
Volume (µL)
Standard
Standard Volume
(µL)
1 RNase P Std 1 RNase P reaction mix 74.25 RNase P Std 1 8.25
2 RNase P Std 2 RNase P reaction mix 74.25 RNase P Std 2 8.25
3 RNase P Std 3 RNase P reaction mix 74.25 RNase P Std 3 8.25
4 RNase P Std 4 RNase P reaction mix 74.25 RNase P Std 4 8.25
5 RNase P Std 5 RNase P reaction mix 74.25 RNase P Std 5 8.25

Applied Biosystems 7500 Fast Specifications

General

Brand	Applied Biosystems
Model	7500 Fast
Category	Laboratory Equipment
Language	English