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, 6 nmol, 15 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