A Haberland, R Dallüge, B Erdmann, S Zaitsev, R Cartier, M Schäfer-Korting, M Böttger
{"title":"Polycation-mediated transfection: how to overcome undesirable side effects of sticky DNA complexes.","authors":"A Haberland, R Dallüge, B Erdmann, S Zaitsev, R Cartier, M Schäfer-Korting, M Böttger","doi":"10.1023/a:1019920516714","DOIUrl":null,"url":null,"abstract":"<p><p>Using polycationic transfection one encounters undesired persistent binding to cells of sticky polycation/DNA complexes. These complexes simulate transfection under conditions where no uptake is expected e.g. at 4 degrees C if the uptake is by endocytosis. To overcome this problem, using H1/DNA complexes, we developed an easy and nontoxic method for removing the sticky complexes not taken up during the transfection phase. The cells are simply washed with isotonic (0.1 M) MgCl2 solution, which enables the complete removal of the complexes by their rapid dissolution.</p>","PeriodicalId":21884,"journal":{"name":"Somatic Cell and Molecular Genetics","volume":"25 5-6","pages":"327-32"},"PeriodicalIF":0.0000,"publicationDate":"1999-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1023/a:1019920516714","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Somatic Cell and Molecular Genetics","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1023/a:1019920516714","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
Abstract
Using polycationic transfection one encounters undesired persistent binding to cells of sticky polycation/DNA complexes. These complexes simulate transfection under conditions where no uptake is expected e.g. at 4 degrees C if the uptake is by endocytosis. To overcome this problem, using H1/DNA complexes, we developed an easy and nontoxic method for removing the sticky complexes not taken up during the transfection phase. The cells are simply washed with isotonic (0.1 M) MgCl2 solution, which enables the complete removal of the complexes by their rapid dissolution.
使用多阳离子转染,会遇到不希望的与粘性多阳离子/DNA复合物的细胞的持续结合。这些复合物模拟转染在没有摄取预期的条件下,例如在4摄氏度,如果摄取是通过内吞作用。为了克服这个问题,利用H1/DNA复合物,我们开发了一种简单无毒的方法来去除转染阶段未被吸收的粘性复合物。细胞简单地用等渗(0.1 M) MgCl2溶液洗涤,这使得它们的快速溶解能够完全去除复合物。