Highly specific qPCR probes contain locked nucleic acids (LNA), which improve performance of RT-qPCR assays, especially in case of A-T rich sequences and SNP analysis. LNAs have restricted conformation of the sugar moiety formed by methylene bridge between oxygen atom at 2’-position and carbon at 4’-position. Such predefined conformation of ribose determines excellent hybridization of oligonucleotides even if they bear only 3-5 LNA residues. qPCR probes can contain up to 6 LNA nucleotides, which increases stability of the duplex with the target DNA (melting temperature (Tm)) (see modification for more information). Highly specific qPCR probes can be the optimal choice for SNP determination or other challenging tasks when increased selectivity is required.
Enhanced sensitivity and accuracy of qPCR
Allows more flexibility as shorter probes can be used
Fluorophore-Quencher pairs for qPCR Dual-labeled Probes*
*For Internal modifications - Quenchers and Fluorophores are incorporated via dT residues. To learn more about modifications follow this link. If you are cannot find modifications of your interest, please contact us info@genterra.ru
Fluorophore
Quencher**
FAM
TAMRA
BHQ1
R6G
SIMA
HEX
R6G
BHQ2
VIC
JOE
HEX
ROX
Cy5
Cy5.5
Cy5.5
BHQ3
0
Locked nucleic acids have restricted conformation of the sugar moiety formed by methylene bridge between oxygen atom at 2’-position and carbon at 4’-position. Such predefined conformation of ribose determines excellent hybridization of oligonucleotides even if they bear only 3-5 LNA residues. LNA determines the ideal conformation for Watson-Crick binding (see modifications). Incorporation of LNA significantly increase melting temperature, so qPCR Probes can be shorter in comparison with standard qPCR probes. Shorter probes are more sensitive, have better quenching and a higher signal-to-noise ratio. Moreover, LNA probes offer an improved ability to distinguish mutations or single nucleotide polymorphisms (SNPs).
Sequence length 5 - 40 bases
Up to 6 LNA per probe are possible
Only DNA bases are allowed at the 5' end
A wide range of dye-quencher combinations available
