What is Stbl3?

What is 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 is Neb stable?

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.

How long are Electrocompetent cells good for?

It is best to pre-aliquot the cells and keep in -80 freezer. As long as cells do not got through thaw and unthaw cycles, they should be able to work for 3 years.

How do you make a competent cell stbl3?

Preparation: 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)….Mark.

How do you read e coli genotype?

Genes: In E. coli, the genotype only includes the genes that carry a loss of function mutation. The gene name is listed as three-letters in lower case and italics (DNA methylase is written as dam). Different genes affecting the same function/pathway are identified with different uppercase italic letters.

What is E coli HB101?

HB101 strain is a hybrid K12 x B bacterium, containing the recA13 mutation that minimizes recombination and aidsin insert stability. In addition, it carries the hsdS20(rB-mB-) restriction minus genotype which prevents cleavage of cloned DNA by endogenous restriction enzymes.

Why is E. coli DH5 alpha used for transformation?

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.

Why do we use DH5 alpha?

DH5 alpha has a recA mutation, so it does no heterologous recombination which ensures a higher insert stability . Additionally, it lacks some endonucleases which might digest the plasmids during the isoation procedure. DH5 alpha is additionally competent for blue-white screening.

Can you heat shock Electrocompetent cells?

There is no way to swap competent cells. Electrocompetent cells are prepared to cope with electrotransformation and chimiocompetent cells are made to be transformed via heat shock.

What is a good transformation efficiency E. coli?

Transformation efficiencies between 10^6 and 10^9 represent the normal range for competent E. coli cells.

How can you increase the transformation efficiency of E coli?

Escherichia coli DH5α cells treated with silver nanoparticles alone resulted in significant increase in transformation efficiency compared to the calcium chloride while using plasmid vectors of different sizes, viz. pUC18, pBR322 and pCAMBIA.