Biotek Synergy H1, Operator'S Manual

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o
Filter-based system, top optics: The Detection Limit must be <=0.160 ng/mL to
pass.
o
Monochromator-based system, top optics: The Detection Limit must be
<=0.160 ng/mL to pass.
Linearity Test
1. Calculate the Mean of the four wells for each concentration in columns 1-5.
2. Perform linear regression using these values as inputs:
Using Sodium Fluorescein
x y
1000 Mean of the 1000 pM wells
500 Mean of the 500 pM wells
250 Mean of the 250 pM wells
125 Mean of the 125 pM wells
62.5 Mean of the 62.5 pM wells
Using Methylumbelliferone
x y
100 Mean of the 100 nM wells
50 Mean of the 50 nM wells
25 Mean of the 25 nM wells
12.5 Mean of the 12.5 nM wells
6.25 Mean of the 6.25 nM wells
3. Calculate the R
2
value; it must be >=0.9500 to pass.
Fluorescence Polarization (FP) Test
1. Using the raw data from the Parallel read:
o
Calculate the Mean Blank (wells A6–H6).
o
Calculate the Signal for each HPR well: Subtract the Mean Blank from its
measurement value.
o
Calculate the Signal for each LPR well: Subtract the Mean Blank from its
measurement value.
2. Using the raw data from the Perpendicular read:
o
Calculate the Mean Blank (wells A6–H6).
o
Calculate the Signal for each HPR well: Subtract the Mean Blank from its
measurement value.
Synergy H1 Operator's Manual
Fluorescence Testing | 99