Biotek Synergy HTX, Operator'S Manual

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Chapter 6: Instrument Qualification
Procedure
1.
Create the Gen5 protocol (see page 129).
2.
If you have not already done so, prepare the test solutions (see page 127).
3.
Refer to the Pipette Map on page 128 and pipette the solutions into a clean, 96-
well solid black plate.
4.
Create a Gen5 experiment based on the
Synergy HTX FI_MUB.prt
protocol
and read the plate.
5.
Calculate and analyze the results as described below.
Results Analysis
Sensitivity Test
1.
Calculate the Mean and Standard Deviation of the 16 reads for each of the
buffer wells (C9, D9, E9).
2.
Among the three buffer wells, find the Median Standard Deviation and
corresponding Mean.
3.
Calculate the Mean for the 16 reads of the MUB Concentration well (D7).
4.
Calculate the Signal-to-Noise Ratio (SNR) using the Mean MUB Concentration
and the Buffer Media STD with its corresponding Buffer Mean:
(MUB Mean –
Buffer Mean)/(3 * Buffer STD)
5.
Calculate the Detection Limit using the known concentration value of MUB and
the calculated SNR:
17.6/SNR
Optic Probe
Detection Limit must
be less than or equal
to:
Top 3 mm 0.16 ng/mL (0.91 nM)
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:
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-Square value; it must be ≥ 0.9500 to pass.
BioTek Instruments, Inc.