pBin19 vector (Cat. No.: V010907)

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Basic Information

Note: pBin19 is a widely used binary vector for Agrobacterium-mediated plant transformation. It features left and right T-DNA borders flanking a cloning site and a kanamycin resistance gene (nptII) for selection in plants. The vector has an RK2 replication origin, allowing it to exist in 10-15 copies per Agrobacterium cell, which can enhance transformation efficiency. Its disarmed T-DNA region enables the insertion of foreign genes to create transgenic plants without causing crown gall disease.

Name:: pBin19

Antibiotic Resistance:: Kanamycin

Length:: 11777 bp

Type:: Plant Vectors

Replication origin:: oriV

Host:: Plants

Source/Author:: Bevan M.

Selection Marker:: Neomycin/G418(Geneticin)

Copy Number:: High copy number

Promoter:: NOS

5' Primer:: M13 fwd

3' Primer:: M13 rev

$ 198.7

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Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Resources

Plasmid Protocol

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5. Store the plasmid at -20 ℃.

6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it

General Plasmid Transform Protocol

1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.

2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.

3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.

4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.

5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.

References

van Engelen FA, Molthoff JW, Conner AJ, Nap JP, Pereira A, Stiekema WJ. pBINPLUS: an improved plant transformation vector based on pBIN19. Transgenic Res. 1995 Jul;4(4):288-90. doi: 10.1007/BF01969123. PMID: 7655517.

Sripriya R, Sangeetha M, Parameswari C, Veluthambi B, Veluthambi K. Improved Agrobacterium-mediated co-transformation and selectable marker elimination in transgenic rice by using a high copy number pBin19-derived binary vector. Plant Sci. 2011 Jun;180(6):766-74. doi: 10.1016/j.plantsci.2011.02.010. Epub 2011 Mar 1. PMID: 21497712.