Identification of the 8-kb plasmid p5343 is described in a previous report (14 (link)). Briefly, the plasmidome of background groundwaters at the ORFRC was examined using an optimized alkaline hydrolysis method followed by Illumina sequencing and analysis. Based on sequence coverage, p5343 was determined to be highly abundant in the background wells studied (14 (link)). In the present study, p5343 was synthesized and cloned into the vector pUC57 using the GenBrick synthesis service (GenScript, Piscataway, NJ) for successful propagation in E. coli . The resulting plasmid p5343_UC57 and its sequence are available via Addgene (Addgene ID 126645) and contain an Escherichia coli compatible origin along with an antibiotic marker for propagation and maintenance in the strain E. coli DH10B. Plasmid purifications were performed using the miniprep plasmid extraction kit (Qiagen GmbH, Germany).
Protocol detail
Isolation and Cloning of Plasmid p5343
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 : Lawrence Berkeley National Laboratory, University of California, Berkeley
Top 5 similar protocols
Variable analysis
independent variables
- Synthesis and cloning of plasmid p5343 into the vector pUC57
- Successful propagation of the resulting plasmid p5343_UC57 in E. coli
- Use of an E. coli compatible origin in the plasmid p5343_UC57
- Inclusion of an antibiotic marker for propagation and maintenance in the E. coli DH10B strain
- Plasmid purification using the miniprep plasmid extraction kit (Qiagen GmbH, Germany)
- No positive or negative controls were explicitly mentioned in the provided information.
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!