NDLI: Nested PCR in magnetically actuated circular closed-loop PCR microchip system

Content Provider	Springer Nature Link
Author	Lok, Khoi Seng Lee, Peter Peng Foo Kwok, Yien Chian Nguyen, Nam Trung
Copyright Year	2012
Abstract	We demonstrate a new and sensitive amplification technique (referred to as Nested Polymerase Chain Reaction; nPCR). It based on a magnetically actuated circular closed-loop PCR microchip system. nPCR involves the use of two sets of primers in two successive PCR runs, and allows the amplification of a single locus from a minute quantity of template DNA. Two sets of primers are specially designed to a target 500-bp region of the bacteriophage lambda template DNA in the first PCR run, and a 247-bp region of the targeted 500-bp first PCR product in the second PCR run. PCR is run on the microchip system and concurrently in regular thermocycler for comparison. The products are analyzed by conventional agarose gel electrophoresis. The detection limit for the initial template DNA is 1.63 × 105 copies per μL (or 8.67 pg) for the first PCR run, and 1.63 copies per μL (or 0.0867 fg) for the second run. The results are comparable to a regular thermocycler. This preliminary study opens a new gateway to future development of specialized nPCR on chip. Figure Nested polymerase chain reaction (nPCR) was first demonstrated on the magnetically actuated circular closed-loop PCR microchip system, using DNA amplification on bacteriophage lambda template DNA as a chemical model.
Starting Page	111
Ending Page	117
Page Count	7
File Format	PDF
ISSN	00263672
Journal	Microchimica Acta
Volume Number	177
Issue Number	1-2
e-ISSN	14365073
Language	English
Publisher	Springer Vienna
Publisher Date	2012-01-19
Publisher Place	Vienna
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Nested PCR Magnetically actuated PCR microchip Lab-on-a-chip Micro total analysis system Nanotechnology Characterization and Evaluation of Materials Nanochemistry Microengineering Analytical Chemistry
Content Type	Text
Resource Type	Article
Subject	Analytical Chemistry

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