School of Minerals Processing and Bioengineering, Central South University, Changsha, China.
Biotechnol Lett. 2011 Aug;33(8):1615-9. doi: 10.1007/s10529-011-0596-6. Epub 2011 Mar 24.
A colony PCR technique was applied for both genomic and chloroplast DNA in the green microalgae Chlorella. Of five different lysis buffers, Chelex-100 was superior for DNA extraction, PCR and DNA storage. It also was insensitive to variations in cell density. The conditions established for an improved PCR formulation are applicable for screening of genetically-engineered transformants as well as bioprospecting of natural microalgal isolates. Besides multiple Chlorella species, we also demonstrate the efficacy of Chelex-100 for colony PCR with a number of other microalgal strains, including Chlamydomonas reinhardtii, Dunaliella salina, Nannochloropsis sp., Coccomyxa sp., and Thalassiosira pseudonana.
一种集落 PCR 技术被应用于绿藻小球藻的基因组和叶绿体 DNA。在五种不同的裂解缓冲液中，Chelex-100 对于 DNA 提取、PCR 和 DNA 储存都是优越的。它对细胞密度的变化也不敏感。为改进 PCR 制剂建立的条件适用于筛选基因工程转化体以及天然微藻分离物的生物勘探。除了多种小球藻物种外，我们还证明 Chelex-100 对其他一些微藻菌株的集落 PCR 也是有效的，包括莱茵衣藻、盐藻、钝顶螺旋藻、寇氏隐甲藻和拟南芥。
