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Nucleic Acid Detection Technology from Olink Bioscience
Padlock probe technology
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| When a Padlock Probe (green, yellow, red strands) hybridizes to the target DNA (blue), its end sequences (red and yellow) can be ligated to form a circular molecule. |
A Padlock probe is a linear oligonucleotide molecule designed with end sequences that are complementary to a sequence in a target DNA molecule. When the end sequences of the Padlock probe hybridize to the target molecule, they are next to each other (in juxtaposition) and can be joined by ligase enzyme to form a circular molecule.
How does a Padlock probe work?
When the Padlock probe is ligated, a circular molecule is formed that can be specifically detected using any DNA amplification method. Since it is circular, the probe is particularly well-suited for rolling circle amplification (RCA). A circularized Padlock probe in combination with rolling circle amplification can be applied to in situ detection, generating a strong localized signal with preserved spatial information in fixed tissue sections or cells. Find out more about this technology applied to protein detection: PLA™ Technology.
The Padlock strategy offers several advantages over other designs. A very large number of probes can be analyzed simultaneously in a sample (high degree of multiplexing) since any cross reactivity between probes will generate linear molecules that will not be amplified and detected. In fact 100,000 Padlock probes have successfully been multiplexed in a single vessel. The result from a Padlock assay can be analyzed in a simple plate reader or in several other formats including microarrays. Padlock probes have successfully been applied to e.g. genotyping, expression profiling, identification of micro-organisms and several other applications.
To read more about rolling circle amplification of Padlock Probes, see Banér et al., Nucleic Acids Research (1998) 26:5073-5078.
The Power behind the Affymetrix Molecular Inversion Probe system
Padlock probe technology lies behind the Affymetrix Molecular Inversion Probe (MIP) system, enabling massively parallel SNP analysis exceeding 100,000 simultaneous SNP typings in one reaction. Find out about this technology at Affymetrix.
To find more information about Padlock Probe technology, visit our publications page.