We maintain two previously characterized strains of C. uncinata, Pol = ATCC PRA-256, USA = USA-SC2, following protocols in Katz et al. (2011) (link). To isolate DNA, cultures were treated overnight with antibiotics and cells were pelleted by spinning at 5,000 rpm for 20 min. Genomic DNA was extracted using phenol/chloroform following standard protocols (Ausubel et al. 1993 ). Micronuclear DNA was isolated according to Katz and Kovner (2010) . Briefly, micronuclear DNA was isolated by gel electrophoresis using Low Melt UltraClean™ Agarose (Mobio15005-50, Carlsbad, CA) after digesting with Bal-31 Nuclease (New England Biolabs M02135, Ispwich, MA) to enrich micronuclear DNA. Gel isolated micronuclear DNA was purified using β-agarase (New England Biolabs M03925).
Protocol detail
Isolation and Purification of Micronuclear DNA from Ciliate Strains
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Corresponding Organization :
Other organizations : Smith College, Ocean University of China, University of Massachusetts Amherst
Top 5 similar protocols
Protocol cited in 1 other protocol
Variable analysis
independent variables
- Strains of C. uncinata: Pol = ATCC PRA-256, USA = USA-SC2
- Genomic DNA
- Micronuclear DNA
- Protocols followed in Katz et al. (2011) for maintaining the two strains
- Overnight treatment with antibiotics
- Pelleting cells by spinning at 5,000 rpm for 20 min
- Genomic DNA extraction using phenol/chloroform following standard protocols (Ausubel et al. 1993)
- Micronuclear DNA isolation according to Katz and Kovner (2010), including Bal-31 Nuclease digestion and gel electrophoresis using Low Melt UltraClean™ Agarose, followed by purification using β-agarase
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!