1997/01/01 by Muqing Cao, Yu Fu, Yan Guo +2
Biochemistry, Genetics and Molecular Biology · Earth and Planetary Sciences · Energy · Social Sciences · #Adult and Continuing Education Topics #Algal biology and biofuel production #Education Systems and Policy #Global Educational Policies and Reforms #Marine and coastal ecosystems #Protist diversity and phylogeny
paper · doi:10.1007/s00709-009-0036-9
openalex publication_date 1997/01/01 · openalex created_date 2016/06/24 · openalex updated_date 2026/07/23
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.