Baier Grit, Musyanovych Anna, Mailänder Volker, Landfester Katharina
Max Planck Institute for Polymer Research, Mainz, Germany.
Int J Artif Organs. 2012 Jan;35(1):77-83. doi: 10.5301/ijao.5000076.
We report the encapsulation of dsDNA molecules with a defined number of base pairs (476 bp and 790 bp) and their subsequent amplification by polymerase chain reaction (PCR) inside nanosized polymeric capsules/droplets. In the first set of experiments, the dsDNA template and PCR reagents were encapsulated in crosslinked potato starch using the inverse (water-in-oil) miniemulsion technique. After redispersion of the capsules in a water-surfactant mixture, PCR was performed inside the crosslinked starch nanocapsules. In the second set of experiments, the PCR was performed inside the aqueous nanodroplets before capsule formation, and then each miniemulsion droplet was covered with a polybutylcyanoacrylate (PBCA) shell which was formed through anionic polymerization directly at the droplet interface. The PCR efficiency was quantitatively evaluated by fluorescence spectroscopy, using a DNA-specific dye called SYBR® Green which intercalates between the base pairs of the dsDNA.
我们报道了对具有特定碱基对数(476个碱基对和790个碱基对)的双链DNA分子进行封装,并随后在纳米级聚合物胶囊/液滴内通过聚合酶链反应(PCR)对其进行扩增。在第一组实验中,使用反相(油包水)微乳液技术将双链DNA模板和PCR试剂封装在交联马铃薯淀粉中。将胶囊重新分散在水 - 表面活性剂混合物中后,在交联淀粉纳米胶囊内进行PCR。在第二组实验中,在胶囊形成之前在水性纳米液滴内进行PCR,然后每个微乳液液滴用聚氰基丙烯酸丁酯(PBCA)壳覆盖,该壳通过阴离子聚合直接在液滴界面形成。使用一种名为SYBR® Green的DNA特异性染料通过荧光光谱法定量评估PCR效率,该染料可插入双链DNA的碱基对之间。