Transposon Sequencing for Bacterial Mutants

For Azoarcus olearius DQS4^T, a bacterial culture at mid-log phase grown at 37°C was collected for DNA extraction. Cells were pelleted by centrifugation and immediately frozen at −80°C until DNA extraction. Genomic DNA was extracted, and transposon insertion junctions were selectively amplified, as described previously (79 (link)). Insertion junction libraries were multiplexed and sequenced using the Illumina HiSeq 2500 system. For Herbaspirillum seropedicae SmR1, libraries were sequenced on either the HiSeq 2000 or HiSeq 2500 system (Illumina) to map a greater fraction of the mutant population. Genomic DNA was extracted, and barcodes were amplified as described previously (16 (link), 18 (link), 80 (link)).

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.

do Amaral F.P., Tuleski T.R., Pankievicz V.C., Melnyk R.A., Arkin A.P., Griffitts J., Tadra-Sfeir M.Z., Maltempi de Souza E., Deutschbauer A., Monteiro R.A, & Stacey G. (2020). Diverse Bacterial Genes Modulate Plant Root Association by Beneficial Bacteria. mBio, 11(6), e03078-20.

Publication 2020

HiSeq 2500 system HiSeq 2000

Azoarcus olearius Bacterial Cells Centrifugation Frozen Genomic Herbaspirillum seropedicae Transposon

Corresponding Organization :

Other organizations : University of Missouri, Universidade Federal do Paraná, Lawrence Berkeley National Laboratory, Brigham Young University

Top 5 similar protocols

Variable analysis

independent variables

Growth temperature: 37°C

dependent variables

Transposon insertion junctions
Fraction of mutant population mapped

control variables

Bacterial culture at mid-log phase

controls

No positive or negative controls were explicitly mentioned.

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.

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?

Revolutionizing how scientists
search and build protocols!