Intact Genomics (ig®) DirectPlate™Competent cells offer simple, fast, and robust results for your DNA transformation needs. DirectPlate™ BL21(DE3) chemically competent E. coli cells are a perfect choice for researchers looking to simplify their transformation workflow by eliminating heat shock, lengthy incubations, and time-consuming outgrowth procedures. Simply mix and directly plate! DirectPlate™ BL21(DE3) chemically competent E. coli cells provide higher transformation efficiency than any competitor‘s similar product and are suitable for high-efficiency transformation in a wide variety of applications such as cloning, protein expression and sub-cloning.
Please find a video below displaying the convenience and capability of this product.
Specifications
Competent cell type: Chemically competent
Species: E. coli Format: Tubes
Transformation efficiency: ≥1 x 107 – 108 cfu/µg pUC19 DNA
Blue/white screening: No
Shipping condition: Dry ice
DirectPlate™BL21(DE3) competent cells: -80 ºC
pUC19 control DNA: -20 ºC
Genotype
F–ompT hsdS(rB– mB–) gal dcm λ(DE3)
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 – 109 CFU/µg pUC19 DNA. Untransformed cells are tested for appropriate antibiotic sensitivity.
General Guidelines
Follow these guidelines when using DirectPlate™ BL21(DE3) 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 gently pipetting up and down a few times.
Comparison Testing
Intact Genomics has performed extensive comparison testing to prove our DirectPlate™ BL21(DE3) chemically competent cells have the highest transformation efficiency on the market.1019-12 1019-36
Use this procedure to transform DirectPlate™ BL21(DE3) 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. No heat shock or lengthy incubations are required.
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 thawed tube of competent cells
3) After adding DNA, mix by gently pipetting up and down a few times then place on ice for 3 minutes.
4) Spread 25 to 50 μl from each transformation directly onto ampicillin 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 a sterilized spreader or autoclaved ColiRoller™ plating beads to spread evenly.
5) Incubate the plates overnight at 37 °C.
Note: The procedures above are for plasmids containing Ampicillin resistant markers
This procedure will increase transformation efficiency nearly 10-fold for DirectPlate™ BL21(DE3)chemically competent cells.
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 thawed tube of competent cells
3) After adding DNA, mix by gently pipetting up and down a few times on ice for ~5 min.
4) Add 950 µl of IG Recovery Media (Cat.# 1711, purchase separately) and shake-incubate at 37 °C, 200rpm for 1 hour
5) Spread 50 to 100 μl from each transformation directly onto antibiotic selection plates (37 °C pre-warmed prior to plating). 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 a sterilized spreader or autoclaved ColiRoller™ plating beads to spread evenly.
6) Incubate the plates overnight at 37 °C.
Note: The procedures above are necessary to obtain high transformation efficiency for plasmids containing chloramphenicol, kanamycin, tetracycline or other resistant markers. For plasmids containing Ampicillin-resistant markers, this procedure will also increase efficiency near 10X compared to the Fast Transformation Protocol.
Manual -ig® DirectPlate™ BL21(DE3) chemically competent E. coli cells
