Molecular cloning, kits: TransformAid™ Bacterial Transformation Kit

TransformAid™ Bacterial Transformation Kit

#K2710

for 20 transformations

#K2711

for 40 transformations

Related Documents (in pdf, ~100 KB):

Detailed Protocol: #K2710, #K2711
MSDS (English)
MSDS (English-USA)
MSDS (German)
Flyer (319 KB)
QuickProtocol™ (55 KB)

Features

Efficient - typical transformation efficiency is more than 10⁷ transformants per µg of plasmid DNA.
Fast - less than one hour from the overnight bacterial culture to the cell plating. Starting with the bacterial colonies, the whole procedure requires only 2.5 hours. No time is wasted on a heat shock or on the incubation of transformed cells in SOC medium before cell plating.
Practical - E.coli competent cells can be prepared from an overnight culture, stored up to 7 days at 4°C, or from bacterial colonies on LB plates stored up to 10 days at 4°C.
Easy - all procedures are performed on ice on your bench. Brief centrifugations are carried out in regular microcentrifuges.
Versatile - suitable with most E.coli strains commonly used for cloning. The kit is also effective with dam- dcm- strains of E.coli.

Description
The TransformAid™ Bacterial Transformation Kit provides a new 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, suitable 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 applied to most E.coli strains commonly used for cloning, including JM107, JM109, XL1-Blue, SURE, TOPP2, W3110, NM527, AD494, CJ236, GM2163, C600, BL21.

Applications

Routine cloning experiments.

Blunt-end cloning.

TA cloning.

Quality Control
The kit is tested in transformation of bacterial strains XL1-Blue and JM107 with pUC19 DNA. Typical transformation efficiency is more than 10⁷ transformants per µg of supercoil pUC19 DNA.

Components of the Kit

C-Medium

T-Solution (A)

T-Solution (B)

Detailed Protocol (in pdf, ~122 KB):

#K2710, #K2711

Note
* Lyophilized bacterial strains JM107 (#M0109) and dam- dcm- GM2163 (#M0099) can be provided free of charge upon request, with any purchase.
Competent cells prepared with TransformAid™ Bacterial Transformation Kit are suitable for direct use only. Freezing down and storage at -70°C is not recommended.

Transformation with the TransformAid™ Bacterial Transformation Kit.

Transformation Protocol from Overnight E.coli Culture (for 2 transformations)

Add 150 µl of the overnight culture to 1.5 ml of pre-warmed C-Medium. Incubate 20 min at 37°C in a shaker.

Prepare T-Solution - mix 250 µl of T-Solution (A) and 250 µl of T-Solution (B). Keep on ice.

Centrifuge refreshed bacterial cells for 1 min in a microcentrifuge to pellet the cells.

Resuspend cells in 300 µl of T-Solution. Incubate 5 min on ice.

Centrifuge for 1 min in a microcentrifuge and resuspend cells in 120 µl of T-Solution. Incubate 5 min on ice.

Add 1 µl of supercoiled DNA (10-100 pg) or up to 5 µl of ligation mixture (10-100ng vector DNA) into new microcentrifuge tubes. Chill 2 min on ice.

Add 50 µl of the prepared cells to each tube containing DNA. Incubate 5 min on ice.Plate immediately on pre-warmed LB-Ampicillin agar plates. Incubate overnight at 37°C.