Greetings for all of scientist using this platform. I have a little problem. Recently I had done reconstruction of my plasmid (Named PDR111, length = 11,8 kb). Transformed culture i named it W1 Transformant. After the transformation being done, I isolated the plasmid with Geneaid Presto Mini Plasmid Kit and i had done electrophoresis after i got the isolated plasmid. The results will be displayed, PDR111 is my plasmid before reconstruction (circular) as a negative control. As you can see, the band from transformed product seems to be nicked or linear. Does its mean that my transformation success? Because my supervisor told me that isolated plasmid from Presto Kit usually circular. Is it possible that my transformation product be nicked/linear plasmid? Please answer me, thank you
So PDR111 is my plasmid before any reconstruction, and W1 Transformant is my plasmid after reconstruction (modification in some nucleotide, but the length is same, about 11,8 kb). After modification of some nucleotide, i had done transformation in E.coli TOP10 and isolated the transformated culture
The type of plasmid obtained from the isolation of a culture can vary depending on the bacterial strain, its environment, and any genetic engineering or manipulation that may have been performed. However, some common types of plasmids that are frequently isolated from bacterial cultures include:
The specific type of plasmid obtained from a culture would depend on factors such as the bacterial strain being studied, its genetic characteristics, and the selective pressures present in its environment. Additionally, researchers may manipulate or engineer plasmids to carry specific genes or traits of interest for experimental purposes.
Jeremy Mullesa I think your plasmid is ok. PDR111 is overloaded, therefore, it appears to run faster and looks smaller/different from your clone. Load equal amounts. To ensure you have the correct fragment cloned, sequence your construct.
Kais Khudhair al Hadrawi you answer is off-topic and simply generated by ChatGPT. The RG community for sure knows how to use this tool.
Thank you so much for the answer. I lack of fund for doing sequence because my lab here doesnt have that facility, so i must pay someone to do sequence for me
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