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EHA105 (pSoup) Competent Cell


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EHA105 (pSoup) Competent Cell

Cat#
ACC-104
Product Name
EHA105 (pSoup) Competent Cell
Product Overview
The genotype of EHA105 (pSoup) Competent Cell is C58 (rifR) Ti pEHA105 (pTiBo542DT-DNA) Succinamopine (pSoup-tetR). EHA105 strain was transformed from EHA101 strain, and the background of the EHA105 strain is C58 type. The nuclear gene of EHA105 contains a screening tag - rifampicin resistance gene (rif). In order to facilitate the transformation, the EHA105 strain carries a amber basic Ti plasmid pEHA105 (pTiBo542DT-DNA) without self-transport function, which contains the vir gene. The vir gene is an essential component for the insertion of T-DNA into the plant genome, and the T-DNA transfer function of the pEHA105 (pTiBo542DT-DNA) plasmid itself is disrupted, but the foreign binary vector T-DNA can be transferred smoothly. Transferring the pSoup plasmid into the EHA105 strain is the EHA105 (pSoup) strain, which helps the pGreen, 62SK, pGs2 series plasmids to replicate in Agrobacterium and helps the EHA105 (pSoup) strain to have tetracycline (tet) resistance.
Applications
EHA105 (pSoup) Competent Cell is suitable for transgenic operations of rice, tobacco and other plants.
Notes
1. The volume of the added plasmid should not be greater than 1/10 of the competent volume; When the plasmid is impure or contaminated with organic substances such as ethanol, the conversion efficiency drops sharply. 2. Gently handle when mixing in the plasmid. If a high concentration of plasmid is converted, the amount of bacteria ultimately used for plating can be reduced accordingly. EHA105 (pSoup) chemically transformed competent cells have tetracycline resistance, but when screening positive clones after transfer into the target plasmid, it is necessary to add antibiotics resistant to the target plasmid without tetracycline. 3. When the density of positive clones on the plate is too large, the positive clones grow slower and the colonies become smaller due to insufficient nutrition. 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. The excessive concentration of rifampicin is not conducive to the growth of Agrobacterium and will reduce its growth rate and transformation efficiency. 5. The purpose of adding rifampicin to the culture medium is to prevent the growth of bacteria and to screen Agrobacterium. According to the resistance of the strain used, the addition of streptomycin or gentamicin can prevent the loss of Ti plasmid, but these antibiotics is not conducive to the transgenic operation of Agrobacterium. Therefore, in general, Agrobacterium is not added with streptomycin or gentamicin, and the probability of loss of Ti plasmid is extremely low (can be ignored).
Storage
Store at - 80 centigrade for 12 months.
Kit Components
EHA105 (pSoup): 100μl/tube * 10 tube/50 tube/100 tube; pGs2(control vector,10ng/μl): 10μl.
Sensitivity
The conversion efficiency of pGs2 (kanamycin resistance) plasmid in EHA105 (pSoup) Competent Cell is >10^4 cfu/μg DNA.
Sample Type
Rice, tobacco and other plants
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