EHA101 electro competent cells (MOFY-0822-FY540)


Cat: MOFY-0822-FY540
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DescriptionThe EHA101 strain is a C58 type background, and the nuclear gene contains a screening tag-rifampicin resistance gene rif. In order to facilitate the transformation operation, this strain carries a nopaline-type Ti plasmid pEHA101 (pTiBo542DT-DNA) without self-transport function, suitable for Transgenic operation in corn, rice, tobacco and other plants.
Gene TypesC58 (rifR) Ti pEHA101 (pTiBo542 D T-DNA) (kanR) Nopaline
Size50 µL/vial
QC resultTransformation efficiency of pK7WGF2 plasmid (size: 11876 bp) detection:>105 cfu/μg DNA; Transformation efficiency of pK7WGF2-ZH plasmid (size: 40 kb) detection:>5×103 cfu/μg DNA
Note1. When the plasmid is added, the volume should not be greater than 1/10 of the competent volume; if the plasmid is impure or there is organic pollution such as ethanol, the transformation efficiency will drop sharply; if the plasmid is doubled, the transformation efficiency will drop by an order of magnitude.
2. Be gentle when mixing plasmids. Transforming high concentrations of plasmids can correspondingly reduce the amount of bacteria that are ultimately used for plating.
3. When the density of positive clones on the plate is too large, due to insufficient nutrition, the growth of positive clones will slow down and the colonies will become smaller. In order to obtain large colonies, the amount of plasmid should be reduced.
4. The concentration of rifampicin should not be higher than 25 μg/ml. Too high concentration of rifampicin is not conducive to the growth of Agrobacterium and will reduce its growth rate and transformation efficiency. The plate used by our company to calculate the transformation efficiency only contains 50 μg/ml kan. If the plate used contains 20 μg/ml rif, the transformation efficiency will be reduced to 1/2.
5. The purpose of adding rifampicin to the medium is to prevent the growth of miscellaneous bacteria and screen Agrobacterium; adding Ti plasmid to screen antibiotics according to the resistance of the strain used can prevent the loss of Ti plasmid, but Ti plasmid screening antibiotics is not conducive to the transgenic operation of Agrobacterium. Therefore, these antibiotics are generally not considered when culturing Agrobacterium, and the probability of Ti plasmid loss is extremely low (can be ignored).

Properties

ShippingDry ice
StorageStore at -80 °C
Handling AdviceImmediately upon receipt, cells were transferred from dry ice to -80°C refrigerator.
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