FRET probes for different instruments and multiplexing
LightCycler probes or FRET probes are hybridization probes based on the fluorescent resonance energy transfer (FRET) technique. They consist of a pair of oligonucleotides (donor and acceptor) each labelled with a different fluorescent dye.
Acceptor
| 5' Modification |
LC610, LC640, Texas Red, ROX, Cyanine5, Cyanine5.5 |
| 3' Modification |
Phosphate, Spacer C3 |
| Int. Modification |
2'-Deoxyinosine, 2'-Deoxyuridine
|
Donor
| 3' Modification |
Fluorescein |
| Int. Modification |
2'-Deoxyinosine, 2'-Deoxyuridine |
Product specifications:
- Delivered quantity: 1, 3, 5 and 10 nmol
- Probe lengths: 15-40 bases (wobbles non-defined ratio possible)
- HPLC purification is included
- TAT of 5 working days
- Delivery format: dried or liquid at selected concentration
- QC report incl. MALDI-TOF MS spectra free of charge
A ready-to-use qPCR probe dilution buffer (10 mM Tris-HCl; 1 mM EDTA; pH 8) is provided along with all LightCycler Probes (1 ml for up to 3 probes).
Principle of FRET probes
FRET probes consist of a donor and an acceptor probe.
Donor and acceptor are designed to hybridise to adjacent regions in close proximity to each other on the target DNA. The acceptor probe is labelled with a fluorescent dye at the 5' end and the donor probe with fluoresceine at the 3' end. Interaction of the two dyes can only occur when both dyes are bound to their target.
During FRET, the fluorescent donor molecule is excited by an external light source which subsequently transfers its energy to the acceptor fluorophore. The excited acceptor emits light of a different (longer) wavelength, which can be detected and measured.