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TransformAid Bacterial Transformation Kit (Thermo Scientific™)

Thermo Scientific TransformAid Bacterial Transformation Kit uses a novel method for rapid preparation of chemically-competent E. coli cells from overnight bacterial cultures or bacterial colonies. The key component of this system is the unique T-solution, which produces competent cells in a few easy steps. The quick and convenient procedure guarantees transformation efficiencies of more than 107 transformants per µg of plasmid DNA, ideal for routine cloning experiments. The option to use bacterial colonies for preparation of competent cells adds flexibility to any cloning experiment.

The TransformAid Bacterial Transformation Kit can be used with most E. coli strains commonly used for cloning: JM107, JM109, XL1-Blue, SURE, TOPP2, W3110, NM527, AD494, CJ236, GM2163, C600, BL21, M15, NM522, HB101, ER2267.

Highlights

• Efficient—more than 107 transformants per µg of plasmid DNA
• Practical—E. coli competent cells can be prepared from an overnight culture or from bacterial colonies
• Fast—less than 1 hour from the overnight bacterial culture to cell plating. Starting from bacterial colonies, the procedure requires only 2.5 hours
• Easy—all procedures are performed on ice on your bench. Brief centrifugations are carried out in regular microcentrifuges

Applications

• Routine cloning experiments
• Blunt-end cloning
• TA cloning

Includes

• C-Medium
• T-Solution (A)
• T-Solution (B)
• Detailed Protocol

Note

Competent cells prepared with TransformAid Bacterial Transformation Kit are suitable for direct-use only. Freezing down and storage at -70°C is not recommended.

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TransformAid Bacterial Transformation Kit

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PIR2 competent cells are recommended for maintaining constructs that express toxic genes or for libraries. Use them for cloning and maintenance of your donor vector construct (or other vector containing the R6K? origin). It contains the wild-type pir gene
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One Shot™ PIR1 Chemically Competent E. coli (Invitrogen™)

The Echo™ Cloning System is a two-vector system that utilizes the Cre-lox site-specific recombination system of bacteriophage P1 (1,2) for rapid cloning into multiple vectors. Your gene of interest is cloned into one of the specially designed donor vectors which can then be recombined with as many different Echo™ acceptor vectors as desired. The donor and acceptor vectors are placed in a buffer that contains Cre recombinase for a 25-minute, in vitro recombination reaction. Cre binds to the lox site present on each vector, brings them together, then cleaves and covalently reattaches the DNA strands. The end result is a fusion of the donor and acceptor vectors forming a single, functional expression vector.
PIR1 competent cells are for cloning and maintenance of your donor vector (i.e. pUniV5/His-TOPO®) construct (or other vector containing the R6K-gamma origin). It contains a mutant allele of the pir gene that maintains the donor vector construct at ~250 copies per cell.