In my case, I have extracted the pGLO plasmid from E. coli HB 101. I carried out restriction digestion and ligation using pGLO as my vector and my target gene as my insert to form a recombinant plasmid. I then inserted my recombinant vector into competent E. coli HB 101 cells for transformation.
Is what I'm doing okay?
Yes this should be fine as long as the HB101 you transformed does not carry any plasmid, they will simply now carry your recombinant plasmid. Note that HB101 is generally suitable for cloning with the downside of being endA+, however, it your intention is expression and protein purification I would use a different strain.
Yes, this is a fine approach if all you want is a strain to carry and replicate your new cloned plasmid. Be sure to send your new construct for sequencing to verify it doesn't have any errors.
What is the purpose of the project?
You need to make sure there are no plasmids in the your bacteria There are protocols for remove plasmids.
Bacteria usually do not accept multiple plasmids at the same time.
I have already seen protocols in this regard on the following site, which I will send you the link, I hope it will be helpful
https://kiagene.ir/store/product-category/%d9%85%d8%ad%db%8c%d8%b7-%da%a9%d8%b4%d8%aa/
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