Intact Genomics (ig®) HB101 chemically competent E. coli cells are suitable for high-efficiency transformation in a wide variety of applications such as cloning and sub-cloning. E. coli HB101 is a K12 x B hybrid strain, containing the recA13 mutation that minimizes recombination and helps insert stability. In addition, it carries the hsdS20(rB-mB-) restriction minus genotype which prevents cleavage of cloned DNA by endogenous restriction enzymes. HB101 strain does not support Alpha-complementation for blue/white screening.
Competent cell type: Chemically competent
Derivative of: HB101
Species: E. coli
Transformation efficiency: ≥1 x 108 cfu/µg pUC19 DNA
Blue/white screening: No
Shipping condition: Dry ice
Reagents Needed for One Reaction
ig® HB101 chemically competent cells: 50 µl
DNA (or pUC19 Control, 10 pg/µl): 1 µl
Recovery medium: 1 ml
ig® HB101 competent cells: -80 ºC
pUC19 control DNA: -20 ºC
Recovery medium: 4 ºC
F- Lambda- araC14 leuB6(Am) DE(gpt-proA)62 lacY1 glnX44(AS) galK2(Oc) recA13 rpsL20(strR) xylA5 mtl-1 thiE1 hsdS20(rB-, mB-)
Transformation efficiency is tested by using the pUC19 control DNA supplied with the kit and using the high-efficiency transformation protocol. Transformation efficiency should be greater than 1×108 CFU/µg pUC19 DNA. Untransformed cells are tested for appropriate antibiotic sensitivity.
Follow these guidelines when using ig® HB101 chemically competent cells.
- Handle competent cells gently as they are highly sensitive to changes in temperature or mechanical lysis caused by pipetting.
- Thaw competent cells on ice, and transform cells immediately following thawing. After adding DNA, mix by tapping the tube gently. Do not mix cells by pipetting or vortexing.