BL21(DE3) pLysS Chemically Competent Cell

Product Overview

Description

 

BL21(DE3) pLysS Chemically Competent Cell

BL21(DE3) pLysS Chemically Competent Cell is specifically designed for chemical transformation of DNA. It is resistant to chloramphenicol(Cam^R) and permits a transformation efficiency of over 10⁷ cfu/μg DNA (tested by pUC19 plasmid DNA). Control Plasmid I (Amp⁺ ) is used to detect cell expression function, the size of expressed protein is about 25 kDa.

 

Cat. No.	CD701
Specification	CD701-02	10×100 μl
	CD701-03	20×100 μl
Storage:	at -70oC for six months
Genotype	F- ompT hsdS(rB-mB- ) gal dcm(DE3) pLysS Camr
Features	Transformation efficiency: >107 cfu/μg (pUC19 DNA).
	CamR
	Contains pLysS plasmid that expresses the T7 lysozyme gene to reduce the background of the target gene expression without disturbing IPTG functioning.
	Suitable for non-toxic and toxic protein expression.
	Control plasmid I (Amp+) is used for detection of expression function of cell. The protein size is about 25 kDa.

 

Procedures

• Thaw a vial of 100 μl BL21(DE3) pLysS Chemically Competent Cell on ice, aliquot 50 μl of the cells into a prechilled 1.5 ml tube, add target DNA into the tube and mix gently. Incubate the cells on ice for 30 minutes.

• Heat-shock the cells for 45 seconds at 42^oC, and then quickly remove the tube from the 42^oC water bath and place them on ice for 2 minutes. Do not shake the tube during this procedure.

• Add 500 μl of sterile SOC medium or LB medium (no antibiotic) into the tube, mix well and shake at 37^oC for 1 hour at 200 rpm to resuscitate cells.

• According to the experiment requirement (plasmid, transformation of recombinant ligation product), add different volumes of transformed cells into corresponding antibiotic-containing LB medium. Spread the transformed cells on selective plate. Invert the plate and incubate at 37^oC overnight.

 

Notes

• High efficiency transformation can be achieved by transforming cells immediately following thawing.

• Avoid repeated thawing.

• Avoid pipetting cells.

• Gentle handling is required for the entire procedure.

 

CITATIONS

Tu W, et al. 2009. Improved production of holotoxin Stx2 with biological activities by using a single-promoter vector and an auto-induction expression system. 67(2):169-74. Protein Expr Purif. IF=1.429. PMID: 19450690.