Plasmid-ID Kit
(Kit #
1000000168
)
Depositing Lab: Philip Poole
The Plasmid-ID Kit allows for the high-throughput assessment of competitive and effective rhizobial strains.
This kit will be sent as bacterial glycerol stocks in 96-well plate format.
Original Publication
Optimizing Rhizobium- legume symbioses by simultaneous measurement of rhizobial competitiveness and N 2 fixation in nodules Mendoza-Suárez MA, Geddes BA, Sánchez-Cañizares C, RamÃrez-González RH, Kirchhelle C, Jorrrin B, Poole PS. Proc Natl Acad Sci U S A. 2020 May 5;117(18):9822-9831. doi: 10.1073/pnas.1921225117. Epub 2020 Apr 21. PubMed PMID: 32317381
Description
The Plasmid-ID kit allows the high-throughput assessment of competitive and effective rhizobial strains.
These plasmids are provided in the strain ST18 (S17 delta pir delta hemA), which requires supplementation with 50 μg/mL of ALA (5-aminolevulinic acid) for growth.
The Plasmid-ID help us to:
- Directly observe plant-bacterial interactions
- Assess competitiveness of strains without reducing their fitness
- Obtain information about their nitrogen fixation rates
- Identify rhizobia directly from individual nodules
This plasmid library has the potential to help pre-evaluate strains as possible inoculants on a selected legume crop in local soils before launching large-scale field use.
A.) Plasmid-ID Library built by Golden Gate cloning. The plasmid library includes 95 individual and unique reporter plasmids, with the following modules: vector; pOGG026, PU module; PsnifH from pOGG043, SC module; sfGFP from pOGG037 and the product from annealing the oligonucleotide was added as a T module (Plasmid ID) as the unique identifier in each case. High-throughput conjugation was made possible with the use of chemically competent E. coli ST18 cells for transformations, which allowed a biparental mating. B.) The Plasmid-ID library allows simultaneous high-throughput measurement of N2 fixation in individual nodules using green fluorescent protein (GFP) and barcode strain identification (Plasmid ID) through next generation sequencing (NGS).
Kit Documentation
Script to identify IDs from sequencing reads available in GitHub: Plasmid-ID
Protocol for high-throughput conjugation from E. coli ST18 into rhizobial strains in 96-well microtiter plates available in the Supplemental Documents of the associated publication
How to Cite this Kit
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which they were created, and include Addgene in the Materials and Methods of your future publications.
For your Materials and Methods section:
“The Plasmid-ID Kit was a gift from Philip Poole (Addgene kit #1000000168).”
For your Reference section:
Optimizing Rhizobium- legume symbioses by simultaneous measurement of rhizobial competitiveness and N 2 fixation in nodules Mendoza-Suárez MA, Geddes BA, Sánchez-Cañizares C, RamÃrez-González RH, Kirchhelle C, Jorrrin B, Poole PS. Proc Natl Acad Sci U S A. 2020 May 5;117(18):9822-9831. doi: 10.1073/pnas.1921225117. Epub 2020 Apr 21. PubMed PMID: 32317381
Plasmid-ID Kit - #1000000168
- Resistance Color Key
Each circle corresponds to a specific antibiotic resistance in the kit plate map wells.
- Inventory
Searchable and sortable table of all plasmids in kit. The Well column lists the plasmid well location in its plate. The Plasmid column links to a plasmid's individual web page.
- Kit Plate Map
96-well plate map for plasmid layout. Hovering over a well reveals the plasmid name, while clicking on a well opens the plasmid page.
Resistance Color Key
| Kanamycin |
Inventory
| Well | Plasmid | Resistance |
|---|---|---|
| A / 1 | pOPS0491 | Kanamycin |
| A / 2 | pOPS0492 | Kanamycin |
| A / 3 | pOPS0493 | Kanamycin |
| A / 4 | pOPS0494 | Kanamycin |
| A / 5 | pOPS0495 | Kanamycin |
| A / 6 | pOPS0496 | Kanamycin |
| A / 7 | pOPS0497 | Kanamycin |
| A / 8 | pOPS0498 | Kanamycin |
| A / 9 | pOPS0499 | Kanamycin |
| A / 10 | pOPS0500 | Kanamycin |
| A / 11 | pOPS0501 | Kanamycin |
| A / 12 | pOPS0502 | Kanamycin |
| B / 1 | pOPS0503 | Kanamycin |
| B / 2 | pOPS0504 | Kanamycin |
| B / 3 | pOPS0505 | Kanamycin |
| B / 4 | pOPS0506 | Kanamycin |
| B / 5 | pOPS0507 | Kanamycin |
| B / 6 | pOPS0508 | Kanamycin |
| B / 7 | pOPS0509 | Kanamycin |
| B / 8 | pOPS0510 | Kanamycin |
| B / 9 | pOPS0511 | Kanamycin |
| B / 10 | pOPS0512 | Kanamycin |
| B / 11 | pOPS0513 | Kanamycin |
| B / 12 | pOPS0514 | Kanamycin |
| C / 1 | pOPS0515 | Kanamycin |
| C / 2 | pOPS0516 | Kanamycin |
| C / 3 | pOPS0517 | Kanamycin |
| C / 4 | pOPS0518 | Kanamycin |
| C / 5 | pOPS0519 | Kanamycin |
| C / 6 | pOPS0520 | Kanamycin |
| C / 7 | pOPS0521 | Kanamycin |
| C / 8 | pOPS0522 | Kanamycin |
| C / 9 | pOPS0523 | Kanamycin |
| C / 10 | pOPS0524 | Kanamycin |
| C / 11 | pOPS0525 | Kanamycin |
| C / 12 | pOPS0526 | Kanamycin |
| D / 1 | pOPS0527 | Kanamycin |
| D / 2 | pOPS0528 | Kanamycin |
| D / 3 | pOPS0529 | Kanamycin |
| D / 4 | pOPS0530 | Kanamycin |
| D / 5 | pOPS0531 | Kanamycin |
| D / 6 | pOPS0532 | Kanamycin |
| D / 7 | pOPS0533 | Kanamycin |
| D / 8 | pOPS0534 | Kanamycin |
| D / 9 | pOPS0535 | Kanamycin |
| D / 10 | pOPS0536 | Kanamycin |
| D / 11 | pOPS0537 | Kanamycin |
| D / 12 | pOPS0538 | Kanamycin |
| E / 1 | pOPS0539 | Kanamycin |
| E / 2 | pOPS0540 | Kanamycin |
| E / 3 | pOPS0541 | Kanamycin |
| E / 4 | pOPS0542 | Kanamycin |
| E / 5 | pOPS0543 | Kanamycin |
| E / 6 | pOPS0544 | Kanamycin |
| E / 7 | pOPS0545 | Kanamycin |
| E / 8 | pOPS0546 | Kanamycin |
| E / 9 | pOPS0547 | Kanamycin |
| E / 10 | pOPS0548 | Kanamycin |
| E / 11 | pOPS0549 | Kanamycin |
| E / 12 | pOPS0550 | Kanamycin |
| F / 1 | pOPS0551 | Kanamycin |
| F / 2 | pOPS0552 | Kanamycin |
| F / 3 | pOPS0553 | Kanamycin |
| F / 4 | pOPS0554 | Kanamycin |
| F / 5 | pOPS0555 | Kanamycin |
| F / 6 | pOPS0556 | Kanamycin |
| F / 7 | pOPS0557 | Kanamycin |
| F / 8 | pOPS0558 | Kanamycin |
| F / 9 | pOPS0559 | Kanamycin |
| F / 10 | pOPS0560 | Kanamycin |
| F / 11 | pOPS0561 | Kanamycin |
| F / 12 | pOPS0562 | Kanamycin |
| G / 1 | pOPS0563 | Kanamycin |
| G / 2 | pOPS0564 | Kanamycin |
| G / 3 | pOPS0565 | Kanamycin |
| G / 4 | pOPS0566 | Kanamycin |
| G / 5 | pOPS0567 | Kanamycin |
| G / 6 | pOPS0568 | Kanamycin |
| G / 7 | pOPS0569 | Kanamycin |
| G / 8 | pOPS0570 | Kanamycin |
| G / 9 | pOPS0571 | Kanamycin |
| G / 10 | pOPS0572 | Kanamycin |
| G / 11 | pOPS0573 | Kanamycin |
| G / 12 | pOPS0574 | Kanamycin |
| H / 1 | pOPS0575 | Kanamycin |
| H / 2 | pOPS0576 | Kanamycin |
| H / 3 | pOPS0577 | Kanamycin |
| H / 4 | pOPS0578 | Kanamycin |
| H / 5 | pOPS0579 | Kanamycin |
| H / 6 | pOPS0580 | Kanamycin |
| H / 7 | pOPS0581 | Kanamycin |
| H / 8 | pOPS0582 | Kanamycin |
| H / 10 | pOPS0584 | Kanamycin |
| H / 11 | pOPS0585 | Kanamycin |
| H / 12 | pOPS0586 | Kanamycin |