Abstract: An analysis method for quantitative PCR experiments including multiple genes of interest, multiple samples and multiple conditions is provided. For calculating the relative expression status of multiple target genes from multiple samples under multiple different conditions, a calculation method based on up to three different parameters is performed. In addition to normalization against one or multiple reference gene(s) and normalization against a reference sample (calibrator), a third normalization step against a base value is performed in a target-, sample- and condition-specific manner.

Abstract: The invention is directed to a system for performing multi-color real time PCT, comprising a flexible real time PCT instrument and a specific composition or reaction mixture for performing multiplex PCR: In particular, the present invention is directed to a composition or reaction mixture which comprises at least 3, preferably 4-5 and most preferably exactly 4 pairs of FRET hybridization probes. Each pair of said hybridization probes consists of a FRET donor probe carrying a FRET donor moiety and a FRET acceptor probe carrying a FRET acceptor moiety having an emission maximum between 550 and 710 nm.

Abstract: The invention is directed to a system for performing multi-color real time PCR, comprising a flexible real time PCR instrument and a specific composition or reaction mixture for performing multiplex PCR: In particular, the present invention is directed to a composition or reaction mixture which comprises at least 3, preferably 4-5 and most preferably exactly 4 pairs of FRET hybridization probes. Each pair of said hybridization probes consists of a FRET donor probe carrying a FRET donor moiety and a FRET acceptor probe carrying a FRET acceptor moiety having an emission maximum between 550 and 710 nm.

Abstract: The present invention is directed to hybridization probes hybridizing adjacently to another at a target nucleic acid sequence, wherein one member of said hybridization probes comprises (i) a nucleotide sequence entity which is substantially complementary to the sequence of the target nucleic acid, (ii) a fluorescent entity being either a FRET donor entity or a FRET acceptor entity, and (iii) a spacer entity connecting the nucleotide sequence entity and the fluorescent entity.

Abstract: The invention is directed to a system for performing multi-color real time PCT, comprising a flexible real time PCT instrument and a specific composition or reaction mixture for performing multiplex PCR: In particular, the present invention is directed to a composition or reaction mixtuer which comprises at least 3, preferably 4-5 and most preferably exactly 4 pairs of FRET hybridization probes. Each pair of said hybridization probes consists of a FRET donor probe carrying a FRET donor moiety and a FRET acceptor probe carrying a FRET acceptor moiety having an emission maximum between 550 and 710 nm.

Abstract: Compositions, reaction mixtures, and kits, as well as methods for their use, comprising a pair of FRET hybridization probes hybridizing adjacently to a target nucleic acid sequence, each hybridization probe comprising (i) a nucleotide sequence entity which is substantially complementary to the sequence of the target nucleic acid, (ii) a fluorescent entity, being either a FRET donor entity or a FRET acceptor entity, and (iii) a spacer entity connecting the nucleotide sequence entity and the fluorescent entity. The intensity of fluorescence emission from the FRET donor entity and from the FRET acceptor entity is not substantially affected by quenching activity of nucleotide residues present in the sequence of the target nucleic acid or in the nucleotide sequence entities of the hybridization probes.