Abstract
A two-step method is reported for preparation of genomic DNA from the model cyanobacterium Synechocystis that can be performed with minimal equipment and reagents in about an hour. High yields of genetic material can be obtained (200–450 ng/μl) with reasonable purity. A further ethanol precipitation step can be included but is not necessary if template is simply required for polymerase chain reaction (PCR) or digestion. This new protocol is helpful for amplification of genes of interest in early-stage research projects and for low throughput screening of transformants. It is more reliable than colony PCR of Synechocystis cultures, and less involved and cheaper than existing clean-DNA preparation methods. It represents an unusually simple and reliable extraction protocol for the growing body of research making use of this cyanobacterium.
Highlights
The increasing interest in cyanobacteria for biotechnology follows their long history as models for the chloroplast (Bryant, 1994)
Numerous methods exist for cyanobacterial genomic DNA extraction which achieve highquality samples suitable for sequencing
Cyanobacterial colony PCR is often refractory, and material cannot be retained for future PCR reactions
Summary
The increasing interest in cyanobacteria for biotechnology follows their long history as models for the chloroplast (Bryant, 1994). An unusually large research base of genomic, biochemical and physiological data mean that cyanobacteria are considered to provide an excellent genetic framework for synthetic biology (Jones et al, 2009; Kehr et al, 2011) and for drug development (Tan, 2007; Singh et al, 2011; Vijaykumar and Menakha, 2015) by virtue of their native anti-cancer and pro-apoptotic compounds, along with their overproduction of phenylpropanoid precursors (Brey et al, 2020; Costa-Rodrigues et al, 2012) Their use in sustainable bioenergy research has been an area of particular activity (Lindberg et al, 2010; Parmar et al, 2011; Wijffels et al, 2013) including production of bioethanol (de Farias Silva and Bertucco, 2016) or hydrogen (Sakurai et al, 2015), and they have been explored as workhorses for bioplastic production (for review, see Katayama et al, 2018)
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