Two methods for quantifying DNA extracted from poly(lactide-co-glycolide) microspheres

被引：38

作者：

Barman, SP ^{[1
]}

Lunsford, L ^{[1
]}

Chambers, P ^{[1
]}

Hedley, ML ^{[1
]}

机构：

[1] ZYCOS Inc, Lexington, MA 02421 USA

来源：

JOURNAL OF CONTROLLED RELEASE | 2000年 / 69卷 / 03期

关键词：

plasmid DNA; encapsulation; PLGA; microspheres; PicoGreen;

D O I：

10.1016/S0168-3659(00)00308-4

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

DNA can be formulated with synthetic polymers such as poly(lactide-co-glycolide) (PLG) to generate microparticles. Researchers have used either UV spectroscopy or fluorometry with PicoGreen(R) dye to quantify PLG-encapsulated DNA. While the sensitivity of DNA detection and quantification by PicoGreen is higher (similar to 12 pg/ml) compared to UV (similar to0.5 mug/ml), each method as an analytical tool has limitations. The premise of this work addresses the usefulness and limitations of each method to determine encapsulation efficiencies in PLG microspheres post-process, and to quantify release of DNA from microspheres during in vitro release experiments. In addition, assay conditions for accurate and reproducible extraction of DNA from PLG microspheres using a biphasic (aqueous/organic) solvent system are described. It was also determined that residual poly(vinyl alcohol) and DNA isoforms (linear, nicked, supercoiled) affected PicoGreen/DNA fluorescence values. (C) 2000 Elsevier Science B.V. All rights reserved.

引用

页码：337 / 344

页数：8

共 15 条

[1] PicoGreen quantitation of DNA: Effective evaluation of samples pre- or post-PCR [J].