tecan INFINITE M1000 PRO, Instructions For Use Manual, Page: 111 / 137

7. Quality Control
2011-09 Instructions for Use for INFINITE M1000 PRO No. 30064852 Rev. No. 1.0 111
7.4.4.2 Detection Limit ATP 384-well Plate
Pipette the reagents into the wells of a Corning 384-well plate (white, flat bottom)
according to the Plate Layout
Material:
ATP Kit SL 144-041, BioThema AB
Corning 384-well plate, flat bottom, white
100 µl p tips
Plate layout:
Pipette 100 µl of the Blank into the wells A1 – D1 and A3 – D10
Pipette 20 µl of ATP standard 10
-7
M into the wells A2 – D2, add 80 µl of ATP
reagent and mix in well (use fresh tip for each well); ATP reagent must NOT
be contaminated with ATP standard!
<
>
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24
A Bx ATP Bx B B B B B B B
B Bx ATP Bx B B B B B B B
C Bx ATP Bx B B B B B B B
D Bx ATP Bx B B B B B B B
E
F
G
H
I
J
K
L
M
N
O
P
ATP….100µl 2*10-8 M ATP; B….100 µl Blank, Bx…100 µl Blank used for
crosstalk check, not included in calculations below.
Measurement Parameters
Before pipetting the plate, prepare instrument for measurement:
Parameters:
Measurement mode:
Luminescence
Integration time: 1000 ms
Plate definition file: COS384fw.pdfx
Part of the plate: A1 – D10
Start measurement immediately after pipetting!
Evaluation:
Calculate the detection limit in fmol/well:
15
B ATP
B
8
1e
1
* 0.0001 *
mean mean
Stdev * 3 * 10 2
well) imit(fmol/ DetectionL
−
−
−
⋅
=
2*10
-8
Concentration of ATP standard [M]
Stdev
B
Standard deviation of Blank (B: A3 – D9)
mean
ATP
Average of wells filled with ATP standard
mean
B
Average of Blank wells (B: A3 – D9)
0.0001 Conversion into mol/well
1/1e
-15
Conversion into fmol/well