An improved colony PCR procedure for genetic screening of <Emphasis Type="Italic">Chlorella</Emphasis> and related microalgae |
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| Authors: | Minxi Wan Julian N Rosenberg Junaid Faruq Michael J Betenbaugh Jinlan Xia |
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| Institution: | (1) School of Minerals Processing and Bioengineering, Central South University, Changsha, 410083, China;(2) Department of Chemical & Biomolecular Engineering, Johns Hopkins University, Baltimore, MD 21218, USA |
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| Abstract: | 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. |
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