120 | Chapter 6: Instrument Qualification
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 SF Concentration well (D7).
4. Calculate the Signal-to-Noise Ratio (SNR) using the Mean SF Concentration
and the Buffer Media STD with its corresponding Buffer Mean:
(Mean SF – Buffer Mean)/(3 * Buffer STD)
5. Calculate the Detection Limit, in pM, using the known concentration value of
SF and the calculated
SNR: 1000/SNR
Optic Probe
Detection Limit must be less than or
equal to:
Bottom 5 mm 30 pM (10 pg/mL)
Top 3 mm 53 pM (20 pg/mL)
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
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
3. Calculate the R-Square value; it must be ≥ 0.9500 to pass.
Troubleshooting
If any tests fail, please try the suggestions below. If the test(s) continue to fail, print the
results and contact BioTek’s Technical Assistance Center.
• Are the solutions fresh? The open buffer and stock solutions should be
discarded after seven days.
• Are the excitation/emission filters clean? Are they in the proper locations and
in the proper orientation in the filter wheels?
BioTek Instruments, Inc.
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