Working concentration (2 Primer, 2 Probes)

Opening Vials Containing Lyophilized Products:

Spin down before opening. The pellet might be attached to the lid.

Hybridization Probes for the LightCycler®

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. Store dark.

Usage with the LightCycler®

To detect sequence variants using the melting curve 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