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Competent Cells

Competent Cells: E. coli & Other Chemically Competent Cells

 E. coli & Other Chemically Competent Cells

 

Intact Genomics offers a broad selection of high-efficiency chemically competent cells for routine and specialized cloning, protein expression, phage display, recombineering, and library construction. Select strains deliver transformation efficiencies of ≥1 × 10¹° cfu/µg DNA. Manufactured under an ISO 13485:2016-certified quality system, our competent cells provide reliable, consistent performance across multiple formats, including tubes, strip tubes, and 96- and 384-well plates. Custom strains, packaging formats, bulk quantities, and OEM solutions are also available.

 

Application

Product Link

Specialization

Efficiency

Cloning

IG 5-alpha Chemically Competent Cells

Routine cloning and sub-cloning, library construction

≥1.0 × 109

IG 10B Chemically Competent Cells

Routine cloning and sub-cloning, library preparation including large DNA >10 kb

≥1.0 × 1010

IG XL1 Blue Max Chemically Competent Cells

Stable plasmid propagation with blue/white screening

≥1.0 × 1010

HB101 Chemically Competent Cells

Stable plasmid maintenance

≥1.0 × 108

JM109(DE3) Chemically Competent Cells

Cloning and T7 protein expression strain

≥1.0 × 108

JM109 Chemically Competent Cells

Routine transformation and cloning

≥1.0 × 108

MG1655 Chemically Competent Cells

"Wildtype" E. coli strain

≥1.0 × 109

Specialized Cloning

IG Stable 2 Chemically Competent Cells

Cloning direct repeat sequences (unstable DNA)

≥1.0 × 109

IG Recombineering Chemically Competent Cells

Homologous recombination, DNA, plasmid, and BAC engineering

≥1.0 × 109

IG ccdB Resist™ Chemically Competent Cells

Propagation of plasmids containing the ccdB gene

≥1.0 × 109

Protein Expression

IG Autoinduction DE3 Chemically Competent Cells

Optimized for autoinduction of protein expression

≥3.0 × 107

BL21(DE3) Chemically Competent Cells

Routine T7 protein expression

≥1.0 × 109

BL21 Chemically Competent Cells

Routine non-T7 protein expression

≥1.0 × 109

BL21(DE3)pLysS Chemically Competent Cells

T7 protein expression with tighter control for toxic proteins

≥3.0 × 107

IG C41(DE3) Chemically Competent Cells

Improved expression of toxic proteins

≥3.0 × 107

IG C41(DE3)pLysS Chemically Competent Cells

Improved expression of toxic proteins with suppressed basal expression

≥1.0 × 107

IG C43(DE3) Chemically Competent Cells

Most tolerant of toxic proteins

≥3.0 × 107

IG C43(DE3)pLysS Chemically Competent Cells

Most tolerant of toxic proteins with suppressed basal expression

≥5.0 × 106

DL39(DE3) Chemically Competent Cells

Precise control over protein synthesis

≥1.0 × 107

Phage Display

ER2738 Phage Display Chemically Competent Cells

Preparation of antibody or peptide phage display libraries

≥1.0 × 1010

Medium

IG Recovery Medium

Superior formulation for competent cell transformations

High-efficiency

IG Autoinduction Medium

Directly promotes high-yield growth of E. coli and high-level expression of T7 promoter-regulated recombinant proteins

High-efficiency

 

 

 

 

MG1655 Chemically Competent Cells

MG1655 Chemically Competent Cells are made from the “wild type” K-12 strain with minimal genetic manipulation.

Intact Genomics

Catalogue No.DescriptionPack SizePriceQty
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IG-1076-12MG1655 Chemically Competent Cells12x50µl £229.00 Quantity
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IG-1076-48MG1655 Chemically Competent Cells12x200µl £413.00 Quantity
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MG1655 Chemically Competent Cells

MG1655 Chemically Competent Cells are made from the “wild type” K-12 strain with minimal genetic manipulation.

 

Product Description

Intact Genomics (ig®) MG1655 Chemically Competent Cells are suitable for routine DNA transformation and other research purposes. MG1655 is the “wild type” K-12 strain with minimal genetic manipulation having only been cured of the temperate bacteriophage lambda and F plasmid by ultraviolet light and acridine orange (1).

Specifications

Competent cell type:  Chemically Competent
Derivative of:  MG1655
Species:  E. coli
Format:  Tubes
Transformation efficiency:  ≥5 x 106 cfu/µg pUC19 DNA
Blue/white screening:  No
Shipping condition:  Dry ice

Reagents Needed for One Reaction

ig® MG1655 chemically competent cells:  50 µl
DNA (or pUC19 Control, 10 pg/µl):  1 µl
Recovery medium:  1 ml

Storage

ig® MG1655 competent cells:   -80 ºC
pUC19 control DNA:  -20 ºC
Recovery medium:  4 ºC

Genotype

F- lambda– ilvG- rfb-50 rph-1

Genomic Features

ig® MG1655 chemically competent cells have the following genomic features:

  • Cured of the temperate bacteriophage lambda and F plasmid
  • A frameshift at the end of rph
  • ilvG- mutation is a frameshift that knocks out acetohydroxy acid synthase II
  • rfb-50 mutation is an IS5 insertion that results in the absence of O-antigen synthesis

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 5×106 CFU/µg pUC19 DNA. Untransformed cells are tested for appropriate antibiotic sensitivity.

General Guidelines

Follow these guidelines when using MG1655 chemically competent E. coli:

  • 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.

Technical Support
Intact Genomics is committed to supporting the worldwide scientific research community by supplying the highest quality reagents. Each new lot of our products is tested to ensure they meet the quality standards and specifications designated for the product. Please follow the instructions carefully and contact us if additional assistance is needed. We appreciate your business and your feedback regarding the performance of our products in your applications.

 

High Efficiency Transformation Protocol

Use this procedure to transform MG1655 chemically competent cells. We recommend verifying the transformation efficiency of the cells using the pUC19 control DNA supplied with the kit. Do not use these cells for electroporation.

1. Remove competent cells from the -80 °C freezer and thaw completely on wet ice (10-15 minutes).
2. Aliquot 1-5 µl (1 pg-100 ng) of DNA to the chilled microcentrifuge tubes on ice.
3. When the cells are thawed, add 50 μl of cells to each DNA tube on ice and mix gently by tapping 4-5 times. For the pUC19 control, add 1 µl of (10 pg/µl) DNA to a chilled microcentrifuge tube, prior to adding 50 µl of Mix well by tapping. Do not pipette up and down or vortex to mix, this can harm cells and decrease transformation efficiency.
4. Incubate the cells with DNA on ice for 30 minutes.
5. After 30-minute ice incubation, heat shock the cells at 42 °C for 45 seconds.
6. Transfer the tubes to ice for 2 minutes.
7. Add 950 µl of Recovery Medium or any other medium of choice to each tube.
8. Incubate tubes at 37 °C for 1 hour at 210 rpm.
9. Spread 50 μl to 200 μl from each transformation on pre-warmed selection plates. We recommend plating two different volumes to ensure that at least one plate will have well-spaced colonies. For the pUC19 control, plate 50 μl on an LB plate containing 100 μg/ml ampicillin. Use sterilized spreader or autoclaved ColiRoller™ plating beads to spread evenly.
10. Incubate the plates overnight at 37 °C.

5 Minute Transformation Protocol

The following procedure results in only ~10% of the transformation efficiency as the protocol listed above.

1. Remove competent cells from the -80 °C freezer and thaw in your hand.
2. Aliquot 1-5 µl (1 pg-100 ng) of DNA to the microcentrifuge tubes. Do not pipette up and down or vortex to mix, this can harm cells and decrease transformation efficiency.
3. Incubate the cells with DNA on ice for 2 minutes.
4. After 2-minute ice incubation, heat shock the cells at 42 °C for 45 seconds.
5. Transfer the tubes to ice for 2 minutes.
6. Add 950 µl of Recovery Medium at room temperature or any other medium of choice to each tube. Immediately spread 50 μl to 200 μl from each transformation on pre-warmed selection plates. We recommend plating two different volumes to ensure that at least one plate will have well-spaced colonies. For the pUC19 control, plate 50 μl on an LB plate containing 100 μg/ml ampicillin. Use sterilized spreader or autoclaved ColiRoller™ plating beads to spread evenly.
7. Incubate the plates overnight at 37 °C.

Downloads

Manual – MG1655 Chemically Competent Cells
MSDS – MG1655 Chemically Competent Cells

 

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200