Trans109 Chemically Competent Cell is specifically designed for chemical transformation of DNA. It permits a transformation efficiency of over 108 cfu/μg DNA (tested by pUC19 plasmid DNA).
endA1 recA1 gyrA96 thi-1 hsdR17 (rk-, mk+) relA1 supE44 D (lac-proAB) [F′traD36 proAB laqIqZΔM15]
• High transformation efficiency: >108 cfu/μg ( pUC19 DNA).
• The lowest homologous recombination favorable for plasmid DNA preparation.
• Routine cloning.
• Blue/white selection.
•Equilibrate a water bath to 42oC.
•Warm a vial of SOC medium or LB medium to room temperature. Warm selective plates at 37oC for 30 minutes.
•Thaw a vial of 100 μl of Trans109 Chemically Competent Cell on ice, aliquot 50 μl of the cells into a prechilled 1.5 ml tube, add target DNA (1 to 5 μl ) into the tube. Do not mix by pipetting up and down. Incubate the cells on ice for 30 minutes.
•Heat-shock the cells for 45 seconds at 42oC without shaking. Immediately transfer the tube to ice. Incubate on ice for 2 minutes without shaking.
•Add 500 μl of prewarmed SOC medium or LB medium (without antibiotic) into the tube, mix well and shake at 37oC for 1 hour at 200 rpm for cell recovery and for the expression of antibiotic resistance.
•Spread 20 to 200 μl from each transformation vial on a prewarmed selective plate. The remaining can be stored at 4oC and plated the next day if needed.
•Invert the plate and incubate at 37oC overnight.
•Select colonies and analyze by restriction enzyme digestion, PCR, or sequencing.C overnight.
•Higher efficiency transformation can be achieved by transforming cells immediately following thawing.
•Avoid repeated thawing.
•Gentle handling is required for the entire procedure.
|Citations & references|
|Production of poly(3-hydroxypropionate) and poly(3-hydroxybutyrate-co-3-hydroxypropionate) from glucose by engineering Escherichia coli||De-Chuan Meng, et al.||2015 Apr||http://www.sciencedirect.com/science/article/pii/S1096717615000415|
|Cloning, expression, and characterization of a novel xylose reductase from Rhizopus oryzae||Journal of Basic Microbiology||1.823||M Zhang, et al.||2015 Feb|
|Down-regulation of a novel ABC transporter gene (Pxwhite) is associated with Cry1Ac resistance in the diamondback moth, Plutella xylostella (L.)||Insect Biochemistry and Molecular Biology||3.42||Zhaojiang Guo，et al .||2015 Jan||http://www.sciencedirect.com/science/article/pii/S0965174815000168|