Diffusion controlled and temperature stable microcapsule reaction compartments for high throughput microcapsule-PCR
2008 (English)In: Advanced Functional Materials, ISSN 1616-301X, E-ISSN 1616-3028, Vol. 18, no 19, 2930-2937 p.Article in journal (Refereed) Published
A novel approach to perform a high number of individual polymerase chain reactions (PCR) in microcapsule reaction compartments, termed “Microcapsule-PCR” was developed. Temperature stable microcapsules with a selective permeable capsule wall were constructed by matrix-assisted layer-by-layer (LbL) Encapsulation technique. During the PCR, small molecular weight building blocks – nucleotides (dNTPs) were supplied externally and diffuse through the permeable capsule wall into the interior, while the resulted high molecular weight PCR products were accumulated within the microcapsule. Microcapsules (∼110.8 µm average diameter) filled with a PCR reaction mixture were constructed by an emulsion technique having a 2% agarose core and a capsule formed by LbL coating with poly(allylamine-hydrochloride) and poly(4-styrene-sulfonate). An encapsulation efficiency of 47% (measured for primer-FITC (22 bases)) and 98% PCR efficiency was achieved. Microcapsules formed by eight layers of polyelectrolyte and subjected to PCR cycling (up to 95 °C) demonstrated good temperature stability without any significantly changes in DNA retention yield and microcapsule morphology. A multiplex Microcapsule-PCR experiment demonstrated that microcapsules are individual compartment and do not exchange templates or primers between microcapsules during PCR cycling.
Place, publisher, year, edition, pages
John Wiley & Sons, 2008. Vol. 18, no 19, 2930-2937 p.
layer-by-layer assembly; microencapsulation; microreactors; polymerization; polyelectrolytes
IdentifiersURN: urn:nbn:se:liu:diva-92065DOI: 10.1002/adfm.200800388OAI: oai:DiVA.org:liu-92065DiVA: diva2:620049