Chlamydomonas (Chlorophyceae) colony PCR
Chlamydomonas (Chlorophyceae) colony PCR
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衣藻 (Chlorophyceae) 菌落 PCR
DOI：
10.1007/s00709-009-0036-9
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发表时间：
2009-03-01
期刊：
影响因子：
2.9
通讯作者：
Pan, Junmin
中科院分区：
文献类型：
--
作者：
Cao, Muqing;Fu, Yu;Pan, Junmin
菌落聚合酶链反应（PCR）的简易性和有效性允许DNA片段的快速扩增和利用遗传操作筛选大量感兴趣的菌落，包括转化体和突变体。在这里，我们评估了衣原体的菌落PCR。用10 mM乙二胺四乙酸（EDTA）或Chelex-100处理单个菌落，所得透明细胞裂解物用于PCR反应。基因组DNA或掺入基因组的质粒DNA被同等地扩增。我们发现Chelex法在某些情况下上级EDTA法。这种菌落PCR技术将绕过分离基因组DNA用于PCR反应的繁琐过程，并将使基因组DNA片段的快速扩增以及转化体的快速大规模筛选成为可能。
The ease and effectiveness of colony polymerase chain reaction (PCR) has allowed rapid amplification of DNA fragments and screening of large number of colonies of interest including transformants and mutants with genetic manipulations. Here, we evaluated colony PCR in Chlamydomonas. Individual colonies were treated with 10 mM ethylenediaminetetraacetic acid (EDTA) or Chelex-100 and the resulting clear cell lysate was used for PCR reaction. Either genomic DNA or plasmid DNA incorporated into the genome was equally amplified. We found that the Chelex method is superior to EDTA method in certain cases. This colony PCR technique will bypass the tedious process of isolating genomic DNA for PCR reaction and will make it possible for rapid amplification of genomic DNA fragments as well as rapid large-scale screening of transformants.