BL21 (DE3) Chemically Competent Cell

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Catalogue No: abx098101
Price: US$768.50
(Size: 1 ml)
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BL21 (DE3) Chemically Competent Cell

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Datasheet SDS
BL21 (DE3) Chemically Competent Cell is designed for chemical transformation of DNA, with a transformation efficiency of over 107 cfu/µg DNA (tested by pUC19 plasmid DNA). The DE3 strain contains the λDE3 lysogen, which carries the gene for T7 RNA polymerase. The control plasmid (Amp+) is used for detection of cell expression functions (protein size: ~25 kDa). Suitable for high expression of non-toxic proteins.

The genotype is: F- ompT hsdS(rB-mB-) gal dcm(DE3).

The 1 ml size consists of 10 × 100 µl Competent Cells, and 20 μl (0.1 ng/μl) Control Plasmid pUC19.

Target BL21 (DE3) Chemically Competent Cell
Storage Store at -70 °C for up to 6 months. Do not store in liquid nitrogen. Avoid repeated freeze/thaw cycles.
Availability Shipped within 10-20 working days.
Note This product is for research use only. This product is shipped with dry ice.
Directions for use

Recommended Protocol:

  1. Equilibrate a water bath to 42 °C. Bring a vial of SOC medium or LB medium to room temperature. Warm selective plates to 37 °C for 30 minutes.
  2. Thaw 100 µl of BL21 (DE3) Chemically Competent Cell on ice. Aliquot 50 µl of cells into a pre-chilled 1.5 ml tube, then add target DNA (1-5 µl). Do not mix by pipetting up and down. Leave on ice for 30 minutes.
  3. Heat-shock the cells for 45 seconds at 42 °C, without shaking. Immediately transfer to the tube to ice. Leave on ice for 2 minutes without shaking.
  4. Add 500 µl of prewarmed SOC medium or LB medium (without antibiotics) into the tube. Mix well and shake at 37 °C for 1 hour at 200 RPM for cell recovery and expression of antibiotic resistance.
  5. Spread 20-200 µl from each transformation vial onto a pre-warmed selective plate. Cells that have not been plated can be stored at 4 °C and plated the next day if required.
  6. Invert the plates and incubate at 37 °C overnight.
  7. Select colonies and analyse by restriction enzyme digestion, PCR or sequencing.

Note:

Higher efficiency transformation can be achieved by transforming cells immediately following thawing. Avoid repeated thawing. All samples and reagents require gentle handling throughout the entire procedure.
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