I have been having trouble with the Agrobacterium, i have transformed competent cells with my plasmid. The plasmid carries kanamycin resistance, GUS and Cas9. The bacteria grow in the kanamycin selective media however when i do PCR for GUS or Cas9 i get no amplification. I have checked the original plasmid i have transferred with the PCR and i get product. So its not the PCR. Also i have ordered some restriction enzymes to check the bacteria in another way but it will take months to arrive. The plasmid minikit i have used was thermo and quiagen so i thought maybe i try another protocol.
Any advice?
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