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. Do not store in liquid nitrogen. Avoid repeated freeze/thaw cycles.
Shelf Life 6 months.
Availability Shipped within 10-20 working days.
Note THIS PRODUCT IS FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC, THERAPEUTIC OR COSMETIC PROCEDURES. NOT FOR HUMAN OR ANIMAL CONSUMPTION.

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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