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A BSA coated NOA81 PCR chip for gene amplification

Author:
Qin, Kuiwei, Lv, Xuefei, Xing, Qiaorui, Li, Rui, Deng, Yulin
Source:
Analytical methods 2016 v.8 no.12 pp. 2584-2591
ISSN:
1759-9679
Subject:
analytical methods, bovine serum albumin, coatings, cobalt, forensic sciences, gene amplification, genes, microsatellite repeats, mutation, organ-on-a-chip, pH, point-of-care testing, polymerase chain reaction, temperature
Abstract:
Norland Optical Adhesive 81 (NOA81) is an excellent material for a microfluidic chip, but this chip has been rarely used in the field of gene amplification due to the inhibition effect on the polymerase chain reaction (PCR). In this study, a NOA81 PCR chip (NP-chip) was developed with a simple, fast fabrication method. To overcome this inhibition effect, a simple bovine serum albumin (BSA) coating method was introduced and the BSA coated pH and concentration were optimized for a better gene amplification effect. The PCR results of HLA-DRB1 indicated that the BSA coating method could greatly improve the PCR efficiency on the NP-chip. Moreover, two kinds of PCRs were performed on the NP-chip to validate the effectiveness of the BSA coating method. (i) Gene mutation of anti EB variable region gene (AEB-HC6-6) was observed under the conditions of cobalt radiation. (ii) Short Tandem Repeats (STRs) PCR was performed on the NP-chip as the multiple-primers PCR. The results showed that the NP-chip with BSA coating was able to successfully realize single and multiple primer gene amplification. With further improvement of the precise temperature control, this kind of NP-chip would be widely applied in gene amplification and promote the development of the miniature gene amplification device in point of care testing (PCOT), forensic detection, etc.
Agid:
6383623