Biotek Synergy HT, Operator'S Manual

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Chapter 4: Instrument Qualification
BioTek Instruments, Inc.
Procedure
1. Create the Gen5 protocols (see page 95) or KC4 protocols (see page 96).
2. Prepare the test solutions (see page 91).
3. Perform the tests using the Bottom optics:
•
Pipette the solutions for the Corners, Sensitivity, and Linearity Tests into a
clean 96-well quartz or glass-bottom microplate (see the map on page 94).
•
Read the plate using the ‘Synergy HT FI_B.prt’ protocol.
4. Perform the tests using the Top optics:
•
Pipette the solutions for the Corners, Sensitivity, and Linearity Tests into a new
96-well solid black or quartz microplate (see the map on page 94).
•
Read the plate using the ‘Synergy HT FI_T.prt’ protocol.
5. Save and/or print the measurement data.
6. Calculate and analyze the results as described below.
Results Analysis
Corners Test
1. Calculate the Mean of the twelve wells containing the 3.3 nM SF test solution
(A1-A3, A10-A12, H1-H3, H10-H12).
2. Calculate the Standard Deviation of the same twelve wells.
3. Calculate the % CV:
(Standard Deviation/Mean) * 100
The % CV must be less than 3.0 to pass.
Sensitivity Test
1. Calculate the Mean and Standard Deviation for the buffer wells (C10-F12).
2. Calculate the Mean for the 1000 pM (1.0 nM) SF solution wells (C1-F1).
3. Calculate the Detection Limit, in pg/mL:
1000 / ((Mean SF - Mean Buffer)/(3 * Standard Deviation Buffer))
Optic Probe Detection Limit must be less than:
Bottom 5 mm 26 pM (10 pg/mL)
Bottom 3 mm 53 pM (20 pg/mL)
Bottom 1.5 mm 106 pM (40 pg/mL)
Top 3 mm 53 pM (20 pg/mL)
Top 1.5 mm 106 pM (40 pg/mL)*
* Typical performance