title: An improved colony PCR procedure for genetic screening of Chlorella and related microalgae.
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.
authors: School of Minerals Processing and Bioengineering, Central South University, Changsha, China. wanminxi@jhu.edu
authors: Minxi
authors: M
authors: Wan
authors: Julian N
authors: JN
authors: Rosenberg
authors: Junaid
authors: J
authors: Faruq
authors: Michael J
authors: MJ
authors: Betenbaugh
authors: Jinlan
authors: J
authors: Xia
mesh_terms: D002708:Chlorella / Q000145:classification / Q000235:genetics*; D002736:Chloroplasts; D004247:DNA / Q000737:chemistry; D004587:Electrophoresis, Agar Gel; D058086:Microalgae / Q000145:classification / Q000235:genetics*; D058889:Molecular Typing / Q000379:methods* / Q000592:standards; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012337:RNA, Ribosomal, 18S / Q000235:genetics
chemical_list: D012337:RNA, Ribosomal, 18S; D004247:DNA