The major advantages of using SPR-based analysis employing optical-based biosensors are (1) analysis is performed in a few minutes, (2) no label is required, and (3) in protocols employing immobilization of molecular probes on the flow cells, the sensor chip can be reused several times. The main limitation is the high cost for most of the commercially available instruments. With respect to the use of (RUfin – RUi) or (RUres – RUi) values, the choice depends on the ability of the employed probes in discriminating target DNA during the association phase of the BIA analysis. In this case, the higher (RUfin – RUi) values are preferably used for analytical determinations, with highly reproducible results (for instance in the detection of GMO or HIV-1 sequences). We cannot generalize on the use of (RUfin – RUi) values for the detection of point mutations; this strongly depends on the length of the probes and the secondary structures of PCR-generated single-stranded target DNAs. In most instances, when (RUfin – RUi) values are not informative for detecting point mutations, (RUres – RUi) values should be taken into consideration.
Surface Plasmon Resonance-Based Biosensor Technology for Real-Time Detection of PCR Products.
FERIOTTO, Giordana;GAMBARI, Roberto
2013
Abstract
The major advantages of using SPR-based analysis employing optical-based biosensors are (1) analysis is performed in a few minutes, (2) no label is required, and (3) in protocols employing immobilization of molecular probes on the flow cells, the sensor chip can be reused several times. The main limitation is the high cost for most of the commercially available instruments. With respect to the use of (RUfin – RUi) or (RUres – RUi) values, the choice depends on the ability of the employed probes in discriminating target DNA during the association phase of the BIA analysis. In this case, the higher (RUfin – RUi) values are preferably used for analytical determinations, with highly reproducible results (for instance in the detection of GMO or HIV-1 sequences). We cannot generalize on the use of (RUfin – RUi) values for the detection of point mutations; this strongly depends on the length of the probes and the secondary structures of PCR-generated single-stranded target DNAs. In most instances, when (RUfin – RUi) values are not informative for detecting point mutations, (RUres – RUi) values should be taken into consideration.I documenti in SFERA sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
