Limits...
Isothermal amplification methods for the detection of nucleic acids in microfluidic devices.

Zanoli LM, Spoto G - Biosensors (Basel) (2012)

Bottom Line: The use of microfluidic devices to miniaturize amplification protocols reduces the required sample volume and the analysis times and offers new possibilities for the process automation and integration in one single device.In contrast, low temperature isothermal amplification methods have no need for thermal cycling thus requiring simplified microfluidic device features.Here, the use of miniaturized analysis systems using isothermal amplification reactions for the nucleic acid amplification will be discussed.

View Article: PubMed Central - PubMed

Affiliation: Istituto Biostrutture e Bioimmagini, CNR, Viale A. Doria 6, Catania, Italy; E-Mail: lzanoli@unict.it.

ABSTRACT
Diagnostic tools for biomolecular detection need to fulfill specific requirements in terms of sensitivity, selectivity and high-throughput in order to widen their applicability and to minimize the cost of the assay. The nucleic acid amplification is a key step in DNA detection assays. It contributes to improving the assay sensitivity by enabling the detection of a limited number of target molecules. The use of microfluidic devices to miniaturize amplification protocols reduces the required sample volume and the analysis times and offers new possibilities for the process automation and integration in one single device. The vast majority of miniaturized systems for nucleic acid analysis exploit the polymerase chain reaction (PCR) amplification method, which requires repeated cycles of three or two temperature-dependent steps during the amplification of the nucleic acid target sequence. In contrast, low temperature isothermal amplification methods have no need for thermal cycling thus requiring simplified microfluidic device features. Here, the use of miniaturized analysis systems using isothermal amplification reactions for the nucleic acid amplification will be discussed.

No MeSH data available.


Principal mechanism for rolling circle amplification (RCA).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4263587&req=5

biosensors-03-00018-f003: Principal mechanism for rolling circle amplification (RCA).

Mentions: The rolling circle amplification (RCA) method exploits the continuous amplification of a circular DNA template by a strand displacing DNA polymerase. The DNA polymerase displaces the synthesized strand and ‘rolls’ on with DNA synthesis. This method operates at a constant temperature and produces a long single-stranded DNA molecule with tandem repeats of the circular template [79]. Both linear and exponential RCAs have been developed. In linear RCA, a small circle sequence is amplified by polymerase extension of a complementary primer, whereas in exponential RCA two primers are used: the second primer hybridizes with the single-stranded DNA product of the first primer and initiates hyper-branching in the DNA replication [80]. A schematic illustration of the principle of rolling-circle amplification is shown in Figure 3.


Isothermal amplification methods for the detection of nucleic acids in microfluidic devices.

Zanoli LM, Spoto G - Biosensors (Basel) (2012)

Principal mechanism for rolling circle amplification (RCA).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263587&req=5

biosensors-03-00018-f003: Principal mechanism for rolling circle amplification (RCA).
Mentions: The rolling circle amplification (RCA) method exploits the continuous amplification of a circular DNA template by a strand displacing DNA polymerase. The DNA polymerase displaces the synthesized strand and ‘rolls’ on with DNA synthesis. This method operates at a constant temperature and produces a long single-stranded DNA molecule with tandem repeats of the circular template [79]. Both linear and exponential RCAs have been developed. In linear RCA, a small circle sequence is amplified by polymerase extension of a complementary primer, whereas in exponential RCA two primers are used: the second primer hybridizes with the single-stranded DNA product of the first primer and initiates hyper-branching in the DNA replication [80]. A schematic illustration of the principle of rolling-circle amplification is shown in Figure 3.

Bottom Line: The use of microfluidic devices to miniaturize amplification protocols reduces the required sample volume and the analysis times and offers new possibilities for the process automation and integration in one single device.In contrast, low temperature isothermal amplification methods have no need for thermal cycling thus requiring simplified microfluidic device features.Here, the use of miniaturized analysis systems using isothermal amplification reactions for the nucleic acid amplification will be discussed.

View Article: PubMed Central - PubMed

Affiliation: Istituto Biostrutture e Bioimmagini, CNR, Viale A. Doria 6, Catania, Italy; E-Mail: lzanoli@unict.it.

ABSTRACT
Diagnostic tools for biomolecular detection need to fulfill specific requirements in terms of sensitivity, selectivity and high-throughput in order to widen their applicability and to minimize the cost of the assay. The nucleic acid amplification is a key step in DNA detection assays. It contributes to improving the assay sensitivity by enabling the detection of a limited number of target molecules. The use of microfluidic devices to miniaturize amplification protocols reduces the required sample volume and the analysis times and offers new possibilities for the process automation and integration in one single device. The vast majority of miniaturized systems for nucleic acid analysis exploit the polymerase chain reaction (PCR) amplification method, which requires repeated cycles of three or two temperature-dependent steps during the amplification of the nucleic acid target sequence. In contrast, low temperature isothermal amplification methods have no need for thermal cycling thus requiring simplified microfluidic device features. Here, the use of miniaturized analysis systems using isothermal amplification reactions for the nucleic acid amplification will be discussed.

No MeSH data available.