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.

Specifications

Competent cell type:            Chemically competentDerivative of:                        HB101Species:                                E. coliFormat:                                 TubesTransformation efficiency:      ≥1 x 10⁸ cfu/µg pUC19 DNABlue/white screening:           NoShipping condition:               Dry ice

Reagents Needed for One Reaction

ig^® HB101 chemically competent cells:   50 µlDNA (or pUC19 Control, 10 pg/µl):            1 µlRecovery medium:                                    1 ml

Storage

ig^® HB101 competent cells:    -80 ºCpUC19 control DNA:                 -20 ºCRecovery medium:                       4 ºC

Genotype

F- Lambda- araC14 leuB6(Am) DE(gpt-proA)62 lacY1 glnX44(AS) galK2(Oc) recA13 rpsL20(strR) xylA5 mtl-1 thiE1 hsdS20(rB-, mB-)

Quality Control

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.

General Guidelines

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.

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