Authors: Muqing Cao Yu Fu Yan Guo Junmin Pan
Publish Date: 2009/02/26
Volume: 235, Issue: 1-4, Pages: 107-110
Abstract
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 Chelex100 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 largescale screening of transformantsWe thank all other members of our laboratory for helpful discussion This work is supported by the National Natural Science Foundation of China 30671090 30771084 National Basic Research Program of China also called 973 program no 2007CB914401
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