AML cells in general don't transfect very well & typically die shortly after. My guess is that you cultured ghost-cells for 21 days.
The transformants are selected for on agar containing an appropriate antibiotic. For example if your recombinant plasmid contains a kanamycin cassette, then only the cells containing the plasmid will grow on an agar plate containing kanamycin. PCR can then be performed on the colonies to ensure they contain your gene of interest on the plasmid.
R-plasmid
TOL plasmid
You can determine if your bacteria contain a plasmid by performing a plasmid extraction followed by gel electrophoresis to visualize the presence of plasmid DNA. Other methods include PCR amplification of plasmid-specific sequences or using molecular biology techniques like restriction enzyme digestion to confirm the presence of a plasmid.
Using the same restriction enzyme ensures that the DNA insert and plasmid have complementary ends and can be ligated together correctly. This helps to ensure that the gene is inserted in the correct orientation and frame, allowing for proper gene expression. Additionally, it helps to prevent self-ligation of the plasmid without the gene of interest.
A plasmid which encodes genes for its own transfer.
Plasmid is extrachromosomal DNA capable of self replication.
A helper plasmid is one that allows for the beginning of replication and transfer of other plasmids from a donor to a recipient. Without a helper plasmid, transposons will not be expressed in the recipient.
You can have a maximum of 8 plasmid slots.
Recombiant DNA
The plasmid is found in prokaryotic cells.
In the production of a recombinant plasmid, the DNA of interest (insert) and the plasmid vector are both cut with restriction enzymes to create compatible ends. These cut fragments are then ligated together using DNA ligase to produce the recombinant plasmid.