Working concentration (2 Primers, 2 Probes)
Opening Vials Containing Lyophilized Products:
Spin down before opening. The pellet might be attached to the lid.
Hybridization Probes for the LightCycler® Instruments
Delivery of HybProbes is in fixed molar amounts, containing 300 pmol, 1 nmol, 3 nmol or 30 nmol.
Do not freeze lyophilized probes. Products are stable at room temperature. Protect from light exposure
Usage with the LightCycler® Instruments
To detect sequence variants using the melting curve option or for quantification use a final probe concentration in the range of 0.15-0.20 µM, corresponding to 3-4 pmol probe per 20 µl reaction. Optimal concentrations have to be established empirically. In some cases a reduction of the fluorescein probe (3FL) has demonstrated to produce better results.
|
HybProbe LC Red (5LC)/ fluorescein |
Concentration in the reaction |
Molar amounts in Master Mix |
|
per reaction |
Vol. per reaction |
|
|
|
32 samples 640µl |
10 samples 200µl |
1 sample 20µl |
Volume of stock solution/reaction |
|
Primer 1 |
0.50 µM |
320 pmol |
100 pmol |
10 pmol |
1 µl/10 µM |
|
Primer 2 |
0.50 µM |
320 pmol |
100 pmol |
10 pmol |
1 µl/10 µM |
|
Probe 5LC |
0.15 µM |
96 pmol |
30 pmol |
3 pmol |
1 µl/3 µM |
|
Probe 3FL |
0.15 µM |
96 pmol |
30 pmol |
3 pmol |
1 µl/3 µM |
|
or |
|
|
|
|
|
|
Probe 3FL |
0.10 µM |
64 pmol |
20 pmol |
2 pmol |
0.7 µl/3 µM |
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