Hi David,

You should be fine using Stbl3. I've used it plenty of times for retroviral transfections and never had an issue. If you're worried about possible degradation of your DNA you could always digest your plasmid with one restriction enzyme to form a linear strand, and run it on an agarose gel to check. If the gel only has one band of the correct size you're good to go, a smear would indicate degradation.

Stbl3 is optimized for lentiviral vectors and gives high yield in minipreps, other competent cells might not produce as much of your vector.

Hope this helps and good luck!

I think you used PB buffer, it should be fine.

hi

You asked both the question and the answer at the same time.

https://www.researchgate.net/post/Which_Ecoli_strain_to_prevent_unexpected_recombination_of_the_plasmid

https://www.researchgate.net/post/Could_we_use_the_DH5alpha_instead_of_Stbl3_for_lentiviral_plasmid_transformation_and_cloning

good luck

Hi David,

I echo what Sarrah said, the Stbl3 strain works well. I used the traditional Qiagen midi or maxi prep kits to purify the plasmids and I never had an issue. You can also use a GFP expressing vector to monitor your plasmid purification, transfection, and viral production procedures.