How do Stbl3 cells grow?
Use glycerol stock or stock of existing competent Stbl2 to streak cells for single colonies on LB plate without antibiotics and grow over night in bacterial incubator (37°C). Store plate at 4°C. Plate should be good for at least 2 weeks.
What is the difference between Stbl2 and Stbl3?
Stbl2 colonies achieved 1-mm diameter in approximately 24 hrs at 30°C. This population of colonies all carried plasmid clones with deletions (33 of 33 colonies analyzed). Stbl3 colonies achieved 1-mm diameter in 18-20 hrs at 30°C.
Why use Stbl3?
When using lentiviral vectors, Stbl3 has advantages. If your lentiviral vector is big, then Stbl3 for sure will be better to use. Stbl3 have higher transformation efficiency with larger plasmids, and also have less mutation and recombination rates.
What is Neb stable bacteria?
NEB Stable enables the isolation of plasmid clones containing repetitive DNA elements: Plasmid pUC-5xREP contains five 32-bp repeats, making it unstable in a recombination-proficient strain.
What is Stbl3 competent cells?
MultiShot FlexPlate Stbl3 chemically competent E. coli cells are cloning competent cells that are packaged in a rack containing 12 strips of 8 wells to increase productivity for medium-throughput bacterial transformations. Unlike TOP10 E.
How do competent cells work?
Whether through electroporation or chemical methods such as calcium chloride, the process of making competent cells creates temporary pores in a cell’s membrane in order for DNA to pass through. This is done by changing the cell in such a way that enables DNA to easily travel through the cell membrane.
Why are lentiviruses used as vectors?
Gene Delivery Using Viral Vectors Lentiviral vectors are a type of retrovirus that can infect both dividing and nondividing cells because their preintegration complex (virus “shell”) can get through the intact membrane of the nucleus of the target cell.
What are Stbl3?
The Stbl3™ E. coli strain is derived from the HB101 E. coli strain and is recommended for use when cloning unstable inserts such as lentiviral DNA containing direct repeats (e.g., Invitrogen® ViraPower™ Lentiviral Expression kits).
What are DH5 cells?
DH5-Alpha Cells are E. coli cells engineered by American biologist Douglas Hanahan to maximize transformation efficiency. They are defined by three mutations: recA1, endA1 which help plasmid insertion and lacZΔM15 which enables blue white screening.