Product Overview
The genotype of E. coli HT115 is F- mcrA mcrB, IN(rrnD-rrnE)1 rnc14::Tn10(DE3 lavUV5::T7 polymerase). HT115 is an Rnase III-deficient E. coli strain with IPTG-inducible T7 Polymerase activity. It is mainly used for RNAi interference test of C. elegans. The strain could grow normally in LB or 2YT medium, and the presence of Tn10 transposons made it tetracycline resistant. The chromosome of this strain integrated the λ phage DE3 region (DE3 region contains T7 phage RNA polymerase, which can induce a large amount of T7RNA polymerase in the presence of IPTG, thus initiating the expression of ds RNA in nematode), and could express both T7RNA polymerase and Escherichia coli RNA polymerase. It can also be used for protein expression of pET series, pGEX, pMAL and other plasmids.
Kit Components
HT115: 100 μL/tube.
Sensitivity
HT115(DE3) competent cells were prepared by a special process, and the transformation efficiency of pUC19 plasmid was 0.5×108 cfu/μg DNA.
Note
1. It is best for receptive cells to melt slowly in ice, and add the target DNA within 8 minutes of insertion into ice. It should not be placed in ice for too long, as long-term storage will reduce the transformation efficiency.
2. The transformation of high concentration of plasmids can correspondingly reduce the amount of bacteria ultimately used for coating.
3. During induction, the IPTG concentration can be selected according to the experiment (0.1-10 mM).
4. During the transformation of nematode RNAi plasmid, the concentration of antibiotics used can be referred to: ampicillin – 50 μg/mL, tetracycline - 12.5 μg/mL.
5. For protein expression, in order to obtain the required amount of protein, the optimal induction time, temperature, and IPTG concentration need to be optimized by experimenters
Storage
Store at -80 °C for 12 months.
Datasheet 
