What is DH5 alpha used for?

What is DH5 alpha used for?

Our DH5α competent E. coli is a versatile strain used for general cloning and sub-cloning applications, and is available in a wide variety of transformation efficiencies.

Is DH5 alpha ampicillin resistance?

Yes, DH5 alpha carries no resistance to antibiotics. It means it should be manipulated very carefully, using stringent asseptic conditions.

Is DH5 Alpha Dam positive?

Yes, DH5a is dam+ (see http://openwetware.org/wiki/E._coli_genotypes) and yes it blocks your BsaBI digestion.

Why are DH5 alpha competent cells?

MAX Efficiency DH5α Competent Cells are a well-known, versatile strain that can be used in many everyday cloning applications. In addition to supporting blue/white screening, recA1 and endA1 mutations in DH5α cells increase insert stability and improve the quality of plasmid DNA prepared from minipreps.

Are DH5 alpha cells DAM+?

Yes, DH5a is dam+ (see http://openwetware.org/wiki/E._coli_genotypes) and yes it blocks your BsaBI digestion. You have to subclone vector into dam- cells.

What is recA1 mutation?

The recA1 mutation is a single point mutation that replaces glycine 160 of the recA polypeptide with an aspartic acid residue. The mutant recA1 protein has a greatly reduced single-stranded DNA-dependent ATPase activity at pH 7.5 compared to the wild-type protein.

Why is DH5 Alpha not used for expression?

Hi Shadariah, DH5alpha is a strain for high yield plasmid DNA production, not proteins. Your plasmid is most likely under the control of an inducible promoter,depending on the system you are using you have to induce protein production by IPTG or use an autoinduction medium.

Can you isolate plasmid from BL21?

BL21 cells are not designed for plasmid purification but protein expression. BL21 lacks in some proteases but has the endonucleases present. You can make mini-preps and try to isolate the plasmid, but there is a risk that your plasmid may be digested.

What is the DH5-Alpha strain?

This strain was developed by D. Hanahan as a cloning strain with multiple mutations that enable high-efficiency transformations (2). The mutations that the DH5-Alpha strain has are: dlacZ Delta M15 Delta(lacZYA-argF) U169 recA1 endA1 hsdR17(rK-mK+) supE44 thi-1 gyrA96 relA1 (2).

Why DH5-Alpha E coli cells?

GoldBio’s DH5-alpha Chemically Competent E. coli cells are suitable for high efficiency transformation in a wide variety of routine applications such as plasmid isolation, cloning, and subcloning.

What is K-12 derived DH5α?

One of the most common laboratory E. coli strains used to maintain and amplify small plasmid DNA is K-12 derived DH5α. Typical DNA assembly and cloning procedures involve as their last step transformation of the constructed plasmid into competent DH5α cells.

What is the shelf life of dhdh5 Alpha?

DH5-alpha Chemically Competent E. coli cells should be stored at -80°C, pUC19 Control DNA should be stored at -20°C and recovery medium should be stored at 4°C immediately upon arrival. When stored under the recommended conditions and handled correctly, these products should be stable for at least 1 year from the date of receipt.