I use the electroporation technique to transfer mcherry and egfp plasmids to salmonella infantis but there is no colony in the final step. is there any way to reach success or changing a step can be helpful?
I will be thankful if somebody can help
Cheers
I am not an expert in this field, but I am very interested and have researched to find an answer. I received some assistance from tlooto.com for this response. Could you please review the response below to see if it is correct?
To improve plasmid transfer to Salmonella Infantis via electroporation, consider the following optimizations: First, ensure that electroporation conditions such as voltage, pulse duration, and cell concentration are optimized. Use freshly prepared, highly competent cells and pure plasmid DNA at an optimal concentration. Pre-chill the cuvettes and cells to enhance efficiency. Additionally, recover cells in SOC medium for at least 1 hour post-electroporation before plating on selective media [1]. If issues persist, verify the antibiotic selection marker's efficacy and ensure the plasmid's origin of replication is compatible with Salmonella Infantis [2]. Adjust these parameters to increase transformation success.
Reference
[1] Yoshida, T., Lai, T. C., Kwon, G., & Sako, K. (2013). pH- and ion-sensitive polymers for drug delivery. Expert Opinion on Drug Delivery, 10, 1497 - 1513.
[2] Varshosaz, J. (2008). pH-sensitive polymers for cytoplasmic drug delivery. Expert Opinion on Therapeutic Patents, 18, 959 - 962.
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