Biotek Synergy H1, Operator'S Manual

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Calculate the Signal for each LPR well: Subtract the Mean Blank from its
measurement value.
3. Calculate the G-Factor for each LPR well:
(Parallel LPR Signal*(1-0.02))/(Perpendicular LPR Signal*(1+0.02))
4. Calculate the Mean G-Factor.
5. Calculate the Polarization value in mP for each HPR well (“PHPR”):
(Parallel HPR Signal-Mean G Factor*Perpendicular HPR Signal)/(Parallel HPR
Mean G Factor*Perpendicular HPR Signal)*1000
6. Calculate the Mean PHPR, in mP.
The Mean PHPR must be >340 mP to pass.
7. Calculate the Polarization value in mP for each LPR well (“PLPR”):
(Parallel LPR Signal-Mean G Factor*Perpendicular LPR Signal)/(Parallel LPR
Mean G Factor*Perpendicular LPR Signal)*1000
8. Calculate the Standard Deviation of the "PLPR", in mP.
The Standard Deviation of the PLPR must be <5 mP to pass.
Time-Resolved Fluorescence (TRF) Test
1. Calculate the Mean and Standard Deviation of the 16 reads for each of the buffer
wells (A6, B6, C6).
2. Among the three buffer wells, find the Median Standard Deviation and
corresponding Mean.
3. Calculate the Mean for the 16 reads of the Eu Concentration well (A8).
4. Calculate the Signal-to-Noise Ratio (SNR) using the Mean Eu Concentration and
Buffer Median STD with its corresponding Buffer Mean:
(Eu Mean - Buffer Mean)/(3 * Buffer STD)
5. Calculate the Detection Limit, in fM: 20000/SNR
The Detection Limit must be <=250 fM to pass.
100 | Chapter 7: Instrument Qualification Process
BioTek Instruments, Inc.