Tyler M Wittkopp
  • Post-Doc, Howard Hughes Medical Institute & Jack H. Skirball Center for Chemical Biology and Proteomics, The Salk Institute for Biological Studies, United States, United States,
Research fields
  • Plant science
Isolation of Genomic DNA from Chlamydomonas reinhardtii
Author:  Tyler M Wittkopp, date: 05/05/2018, view: 13415, Q&A: 0
Chlamydomonas reinhardtii is a soil-dwelling eukaryotic green alga that is widely used as a laboratory model organism for research on photosynthesis, ciliary biology, lipid metabolism and many other aspects of cell biology and physiology. With sequenced nuclear, chloroplast and mitochondrial genomes, Chlamydomonas is also an excellent organism for genetics and genomics research. This protocol describes the isolation of genomic DNA from Chlamydomonas using a standard phenol:chloroform extraction method followed by ethanol precipitation. The protocol requires minimal lab materials, takes approximately 4 h to complete, and can also be used for isolation of genomic DNA from other eukaryotic green algae.
Nuclear Transformation of Chlamydomonas reinhardtii by Electroporation
Author:  Tyler M Wittkopp, date: 05/05/2018, view: 9657, Q&A: 1
The unicellular green alga Chlamydomonas reinhardtii is an important model organism for studying photosynthesis, acclimation to abiotic stress, cilia biology, and many other biological processes. Many molecular biology tools exist for interrogating gene function including the ability to easily transform the nuclear genome of Chlamydomonas. While technical advances such as TALENs, ZFNs and CRISPR are making it easier to precisely edit the nuclear genome, the efficiency of such methods in Chlamydomonas is at present very low. In contrast, random insertion by nuclear transformation tends to be a much more efficient process. This protocol describes a method for transformation of the Chlamydomonas nuclear genome by electroporation. The protocol requires at least 3 days of work and generally results in the appearance of small colonies within 1-2 weeks.
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