Skip to main content
This website uses cookies to ensure you get the best experience. By continuing to use this site, you agree to the use of cookies.

Please note: Your browser does not support the features used on Addgene's website. You may not be able to create an account or request plasmids through this website until you upgrade your browser. Learn more

Please note: Your browser does not fully support some of the features used on Addgene's website. If you run into any problems registering, depositing, or ordering please contact us at [email protected] Learn more

plasmid kit icon

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.

Add to Cart
$350 USD + shipping

Available to academics and nonprofits only.

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, Ramírez-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.

Plasmid-ID.jpg

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, Ramírez-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
Data calculated @ 2021-07-25

Kit Plate Map - #1000000168

Content blocked, you may need to disable your ad-blocker.