Bio-Rad
Gene Pulser Xcell™ Total System
1652660
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Gram negative bacteria like Salmonella Typhimurium are difficult transform with expression plasmid vectors by chemical method. In such cases, an electro-transformation using Bio-Rad Gene Pulser Xcell system gives higher transformation efficiency and also with difficult transform strains.
Transformation of Salmonella strains with expression plasmids
200 ng of expression plasmid pET30 and electro-competent Salmonella Typhimurium
1. Thaw electro-competent Salmonella Typhimurium cells on ice for 20 min. Then add 200 ng total expression plasmid into the cells and mix by gentle flicking of tube. 2. Transfer above mix (ideally 100 microliter volume) into ice cold 0.2 cm electrode gap cuvette. 3. Place the cuvette in cuvette holder apparatus and perform electro transformation at 2500 volts, 25 microF, 200 ohms.4. Ideal transformation results in delivery of around 2400 to 2500 volts within 5 to 6 milliseconds.5. Immediately recover the cells from cuvette by adding 900 microliter SOC medium and proceed for outgrowth for 2 hr at required temperature and selection suitable selective agar plate.
Avoid air bubbles while transferring competent cells containing expression plasmid. Make sure you cool elctroporation cuvette on ice before performing electro-transformation.
Using Bio-Rad Gene Pulser Xcell apparatus, the transformation efficiency of difficult to transform Salmonella strain was achieved at 10^6 cfu/microgram of plasmid vector.
Ideal apparatus for molecular biology lab. Rarely experience electric arching.
N/A
Easy to set up and program required electro transformation parameters.
None
Highly recommended and must have apparatus for molecular cloning and biotechnology laboratories.