Fast magnetic isolation of simple sequence repeat markers in microfluidic channels.
Lab Chip
; 14(8): 1410-4, 2014 Apr 21.
Article
em En
| MEDLINE
| ID: mdl-24615343
ABSTRACT
Simple sequence repeat (SSR) markers are widely used for genome mapping, genetic diversity characterization and medical diagnosis. The fast isolation by AFLP of sequence containing repeats (FIASCO) is a powerful method for SSR marker isolation, but it is laborious, costly, and time consuming and requires multiple rounds of washing. Here, we report a superparamagnetic bead (SPMB)-based FIASCO method in a magnetic field controllable microfluidic chip (MFCM-Chip). This method dramatically reduces the assay time by 4.25-fold and reduces the quantity of magnetic beads and probes by 10-fold through the magnetic capture of (AG)n-containing fragments from Herba Leonuri, followed by washing and eluting on a microchip. The feasibility of this method was further evaluated by PCR and sequencing, and the results showed that the proportion of fragments containing SSRs was 89%, confirming that this platform is a fast and efficient method for SSR marker isolation. This cost-effective platform will make the powerful FIASCO technique more accessible for routine use with a wide variety of materials.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Repetições de Microssatélites
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Técnicas Analíticas Microfluídicas
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Imãs
Idioma:
En
Ano de publicação:
2014
Tipo de documento:
Article