DNA purification on a lab-on-valve system incorporating a renewable microcolumn with in situ monitoring by laser-induced fluorescence.
Bead injection in a lab-on-valve (LOV) system was adopted for DNA purification via micro solid-phase extraction (SPE) with a renewable silica microcolumn packed in a channel of the LOV unit. The complex matrix components in human whole blood, including proteins, were well eliminated by choosing properly the sample loading and elution media. The DNA purification process was monitored on-line by using laser-induced fluorescence in a demountable side part of the LOV unit incorporating optical fibers. The practical applicability of the entire system was demonstrated by separation/purification of lambda-DNA in a simulated matrix and human blood genetic DNA by performing SPE, in situ monitoring of the purified products, and postcolumn PCR amplification. When DNAs in a simulated matrix (10.0 ng microl-1 lambda-DNA, 50 ng microl-1 bovine serum albumin, 1.0% Triton X-100) were processed in the present system and laser-induced fluorescence was monitored at 610 nm, an overall extraction/collection efficiency of 70% was achieved by employing identical sample loading and an elution flow rate of 0.5 microl s-1, along with a precision of 3.8% relative standard deviation. DNA separation and purification from human whole-blood samples were performed under similar conditions.
Chen
Xu-Wei
XW
Research Center for Analytical Sciences, Northeastern University, Box 332, Shenyang, 110004, China.
Xu
Zhang-Run
ZR
Qu
Bai-Yan
BY
Wu
Ye-Feng
YF
Zhou
Jing
J
Zhang
Hui-Dan
HD
Fang
Jin
J
Wang
Jian-Hua
JH
D004247:DNA / Q000097:blood* / Q000302:isolation & purification*; D017022:Flow Injection Analysis / Q000295:instrumentation / Q000379:methods; D005453:Fluorescence; D006801:Humans; D007834:Lasers; D016133:Polymerase Chain Reaction; D012822:Silicon Dioxide / Q000737:chemistry; D052617:Solid Phase Microextraction; D013050:Spectrometry, Fluorescence
D012822:Silicon Dioxide; D004247:DNA