Trans10 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). The competent cell is resistant to streptomycin sulfate (StrR).
F- mcrA Δ(mrr-hsdRMS-mcrBC) φ80 lacZΔM15Δ lacX74 recA1 araΔ139Δ(ara-leu)7697 galU galK rpsL (StrR) endA1 nupG
• High transformation efficiency: >108 cfu/μg ( pUC19 DNA).
• Blue/white selection.
• Toxic gene cloning and stable replication of plasmid DNA.
•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 Trans10 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 30 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|
|Development and characterization of microsatellite loci for lotus (Nelumbo nucifera)||Conservation Genetics||2.183||Tian H L, et al.||2008 Oct||http://link.springer.com/article/10.1007/s10592-007-9503-z|
|MAPK signaling pathway alters expression of midgut ALP and ABCC genes and causes resistance to Bacillus thuringiensis Cry1Ac toxin in diamondback moth||PLoS Genetics||8.167||Zhaojiang Guo||2015 Apr||http://journals.plos.org/plosgenetics/article?id=10.1371/journal.pgen.1005124|
|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||2015 Jan||http://www.sciencedirect.com/science/article/pii/S0965174815000168|
|Producing aglycons of ginsenosides in bakers' yeast||Scientific Reports||2.927||Zhubo Dai, et al.||2014 Jan||http://link.springer.com/article/10.1007/s12010-015-1759-z|
|Heterologous Expression and Characterization of a GH3 β-Glucosidase from Thermophilic Fungi Myceliophthora thermophila in Pichia pastoris||Applied Biochemistry and Biotechnology||1.735||Junqi Zhao, et al.||2015 Sep||http://www.ncbi.nlm.nih.gov/pubmed/23762300|