Instrument
The capillary-based LightCycler® Instruments from Roche Diagnostics are the fastest thermocyclers with online monitoring of DNA amplification. The system works with heated air, allowing rapid ramping – 30 cycles can be performed in 20 minutes. The LightCycler® Real-Time PCR thermocyclers detect fluorescence in up to six separate channels. For quantification the growing PCR product is monitored using a DNA-binding dye, or with TaqMan® or Hybridization Probes. Some of the major advantages of Real-Time detection are the direct quantification of the target and, by avoiding post – PCR handling, a minimal contamination risk with PCR products.
Genotyping
The LightCycler® Instruments melting curve analysis offers an accurate detection of single base mismatches. This unique feature enables rapid analysis of polymorphisms in genetics, clinical chemistry, forensic analysis, pharmacology and other research fields.
Quantification
When using external standards of known copy numbers, the LightCycler® Instruments will enable the exact determination of initial template amount. When working with plasmid standards, we recommend the use of dUracil-containing preparations to minimize the contamination risk. External custom standards are available from GenExpress GmbH, Berlin.
The LightCycler® instruments differ in the availability of channels for the use of Hybridization Probes.
|
Color (Channels) |
530 |
610 |
640 |
670 |
705 |
|
LightCycler® 1.2 |
F1 |
|
F2 |
|
F3 |
|
LightCycler® 1.5 |
F1 |
|
F2 |
|
F3 |
|
LightCycler® 2.0 |
X |
X |
X |
X |
X |
|
LightCycler® 480 |
X |
X |
X |
X |
|
|
LightCycler® 480 II |
X |
X |
X |
X |
X |
|
LightCycler® Nano |
|
|
|
|
|
Only Channel F2 (640) is available for all systems. For Genotyping duplex testings we use channels F1 (530) and F2 (640); a specific color compensation for calibration of the instrument is a prerequisite. We prefer to use channels F2 (640) and F3 (690) for our quantification in order to be able to work with all (except for LC480 I) LightCycler® instruments.
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