ig® Thymidine-Auxotrophic EHA101 Agro cells includes modifications so that they will not grow unless thymidine is added to Minimal medium. This prevents the bacteria from overgrowing plant tissues.
Thymidine Auxotrophic EHA101Thy Agrobacterium Electrocompetent Cells
$505.00 – $1,100.00
Description
Intact Genomics (ig®) Thymidine-Auxotrophic EHA101Thy Agrobacterium ElectroCompetent Cells include modifications so that they will not grow unless 50 mg/L of thymidine is added to Minimal medium. This prevents the bacteria from overgrowing plant tissues when used for plant transformation. Thymidine-Auxotrophic EHA101Thy Agrobacterium ElectroCompetent Cells are optimized for the highest transformation efficiencies. The EHA101Thy strain is useful for transgenic operations of rice, tobacco and other plants. EHA101Thy– thyA knockout mutant, spectinomycin-resistant, derived from EHA101, itself a derivative of A281 (A136/pTiBo542).
Custom Aliquots Available
Benefits:
- Thymidine Auxotrophic
- Enables development of more efficient transformation systems
- Reduced bacterial overgrowth during co-cultivation
- Decreased need for antibiotics
Specifications:
Competent cell type: Electroporation Competent
Species: A. tumefaciens
Strain: EHA101Thy
Format: Tubes
Transformation efficiency: ≥ 1 x 103 cfu/µg pTF102DNA
Shipping condition: Dry ice
Product Components:
- ig® Thymidine-Auxotrophic EHA101Thy Agrobacterium ElectroCompetent Cells
- DNA (pTF102, 500 pg/µl)
- Recovery medium*
* Client needs to add 50 mg/L of thymidine (no provided) to recovery media before use
Storage:
- ig® EHA101Thy Agrobacterium ElectroCompetent Cells: -80 °C
- pTF102 control DNA: -20 °C
- Recovery medium: 4 °C
1402 1402-10 1402-30
Additional information
| μl | 5×50μl, 15×50μl, 6×100μl, 12×100μl |
|---|
References
Aliu E., Azanu MK., Wang K., Lee K. Generation of thymidine auxotrophic Agrobacterium tumefaciens strains for plant transformation https://www.biorxiv.org/content/10.1101/2020.08.21.261941v1.full.pdf
Additional Information:
The auxotrophic Agrobacterium tumefaciens strains were developed with support by National Science Foundation Plant Genome Research Program Grant 1725122 and 1917138 to K.W., by the Iowa State University Interdepartmental Plant Biology Major fellowship to EA and MA, by the USDA NIFA Hatch project #IOW04341 and by State of Iowa funds and by the Crop Bioengineering Center of Iowa State University.
Technical Support
Intact Genomics (IG®) is dedicated to customer satisfaction regarding the use of our products for your research needs. Each new lot of our products is thoroughly tested to ensure it meets the high quality standards and provide excellent results. We appreciate your business and your feedback regarding the performance of our products in your applications. Please follow the instructions carefully and contact us if additional assistance is needed.
IG Technical Support
Email: [email protected]
Phone: (314) 942-3655 | Toll free: 855-835-7172
Legal Notices
Intact Genomics owns the following registered trademarks granted by the United States Patent and Trademark Office (USPTO): Intact Genomics®, IG®, ig®, igTherapeutics®, FastAmp®, i7®, DirectPlate® .
All technology protocols discussed within this manual are assumed proprietary to Intact Genomics. This Product may be covered by pending or issued patents or may have certain limitations. Please contact us for more information. Purchase of this material conveys to buyer the non-transferable right to use the material purchased in research conducted by buyer, whether for teaching, non-commercial or commercial research purposes. Buyer may not sell or otherwise transfer these materials, its components, or unmodified descendants to a third party.
Product Use Limitation and Disclaimers
This product is intended for research purposes only, not for therapeutic or diagnostic purposes in humans or animals. This product contains chemicals which may be harmful if misused or making direct human contact.
Intact Genomics is dedicated to practicing and maintaining science and technology ethics. Buyer agrees to use the purchased materials in full compliance with applicable law and regulations.
Transformation Protocol:
Use this procedure to transform ig® Thymidine-Auxotrophic EHA101Thy Agrobacterium ElectroCompetent Cells. Do not use these cells for chemically transformation.
- Place sterile cuvettes and microcentrifuge tubes on ice.
- Remove competent cells from the -80 °C freezer and thaw completely on wet ice (10-15 minutes).
- Aliquot 1 µl (10pg -1 µg) of DNA to the chilled microcentrifuge tubes on ice.
- When the cells are thawed, add 25 μl of cells to each DNA tube on ice and mix gently by tapping 4-5 times.
For the pRF102 control, add 1 µl of (500 pg/µl) DNA to the 25 µl of cells on ice. Mix well by tapping.
Do not pipette up and down or vortex to mix, this can harm cells and decrease transformation efficiency. - Pipette 26 µl of the cell/DNA mixture into a chilled electroporation cuvette without introducing bubbles. Quickly flick the cuvette downward with your wrist to deposit the cells across the bottom of the well and
then electroporate. - Immediately add 974 µl of Recovery Medium with 50 mg/L of thymidine or any other medium of choice to the cuvette, pipette up and down three times to re-suspend the cells. Transfer the cells and Recovery Medium to a 15ml of culture tube.
- Incubate tubes at 30 °C for 3 hours at 200 RPM.
- Spread 20-200 μl cells onto a pre-warmed selective plate. For the pTF102 control DNA, you may plate 100 μl of undiluted transformation mix onto a YT plate containing 50 μg/ml of thymidine, 15 μg/ml rifampicin and 50 μg/ml spectinomycin. Use sterilized spreader or autoclaved ColiRoller™ plating beads to spread evenly.
- Incubate the plates for 2 – 3 days at 30 °C.
Electroporation Settings:
Mode: Exponential protocol
Voltage (V): 1,800 V
Capacitance: 25 uFD
Resistance: 200 Ohms
Cuvette: 1 mm
