A polymer lab-on-a-chip for genetic analysis using the arrayed primer extension on microarray chips.
In this work a polymer lab-on-a-chip (LOC), fabricated through MEMS technology, was employed to execute a genetic protocol for the Single Nucleotide Polymorphisms (SNPs) detection. The LOC was made in Poly (methyl methacrylate) (PMMA) and has two levels: the lower one for the insertion and mixing of the reagents, the upper one for the interfacing with the DNA microarray chip. The hereditary hearing loss was chosen as case of study, since the demand for testing such a particular disorder is high and genetics behind the condition is quite clear. The Arrayed Primer EXtension (APEX) genetic protocol was implemented on the LOC to analyze the SNPs. A low density (for detection of up to 10 mutations) and a high density microarray chips (for detection of 245 mutations in 12 genes), containing the primers for the extension, were employed to carry out the APEX reaction on the LOC. Both the microarray chips provide a signal to noise ratio and efficiency comparable with a detection obtained in a conventional protocol in standard conditions. Moreover, significant reduction of the employed PCR volume (from 30 μL to 10 μL) was obtained using the low density chip.
Xlab - Materials and Microsystems Laboratory, Applied Science and Technology Department (DISAT), Polytechnic of Turin - Latemar Unit, Via Lungo Piazza d'Armi 6, 10034, Chivasso (Turin), Italy, simone.marasso@polito.it.
Simone L
SL
Marasso
Domenico
D
Mombello
Matteo
M
Cocuzza
Davide
D
Casalena
Ivan
I
Ferrante
Alessandro
A
Nesca
Piret
P
Poiklik
Kadri
K
Rekker
Anu
A
Aaspollu
Sergio
S
Ferrero
Candido F
CF
Pirri
D017931:DNA Primers / Q000737:chemistry*; D006801:Humans; D056656:Lab-On-A-Chip Devices; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation* / Q000379:methods*; D020641:Polymorphism, Single Nucleotide
D017931:DNA Primers