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Xl1 blue electrocompetent cells

Manufactured by Agilent Technologies
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XL1 Blue Electrocompetent cells are a laboratory product designed for the transformation of DNA into bacterial cells. They facilitate the introduction of plasmids or other genetic material into Escherichia coli cells through electroporation, a process that uses an electric field to increase the cells' permeability and allow the uptake of foreign DNA.

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Lab products found in correlation

Lb broth base, by Thermo Fisher Scientific (1 mentions) Purelink hipure expi plasmid megaprep kit, by Thermo Fisher Scientific (1 mentions) Mouse igg library primer set, by Progen Biotechnik (1 mentions)

Close spelling variants of this product

XL1-Blue

2 protocols using xl1 blue electrocompetent cells

1

Cloning and Expression of MUC2-C Protein

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The pcDNA3.1(+) plasmid was used as a template for inserting the MUC2-C (NCBI reference: MH593786.18 (link) (protein AZL49145.1) containing the amino acids 4356–5130 with an N-terminal IGk signal sequence, a 6 His-tag and the 3C precision cleavage tag in that order from the N-terminal. The mammalian expression vector was transformed through electroporation using XL1 Blue Electrocompetent cells (Agilent). The transformed cells were selected with the corresponding antibiotic and grown overnight in LB Broth Base (Invitrogen). DNA was prepared using PureLink™ HiPure Expi Plasmid Megaprep Kit (Thermo Fisher Scientific).
Gallego P., Garcia-Bonete M.J., Trillo-Muyo S., Recktenwald C.V., Johansson M.E, & Hansson G.C. (2023). The intestinal MUC2 mucin C-terminus is stabilized by an extra disulfide bond in comparison to von Willebrand factor and other gel-forming mucins. Nature Communications, 14, 1969.
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2

Cloning and Expression of Anti-CD19 scFv Antibody

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The nucleotide sequences of the cloned variable heavy (VH) and variable light (VL) domains of the anticanine CD19 antibody (clone 4E9) were amplified from hybridoma cDNA using the Mouse IgG Library Primer Set (Progen, Heidelberg, Germany). The variable domains were synthesized into scFv formats in the VH-VL and VL-VH orientation using the Whitlow peptide linker49 (link) and cloned into the pComb3X phagemid vector (a kind gift from Dr. Donald Siegel). XL-1 BLUE electrocompetent cells (Agilent Technologies, Santa Clara, CA) were transformed and cultured in the presence of carbenicillin to select for transformed colonies. Phagemid was rescued with the addition of M13 helper phage and further selected using kanamycin. Resulting phage was precipitated with 5× PEG/NaCl and resuspended in 1% BSA in PBS. The titer of each phage preparation was determined using a plaqueforming assay as previously described.3
Preethi Haran K., Lockhart A., Xiong A., Radaelli E., Savickas P.J., Posey A, & Mason N.J. (2020). Generation and Validation of an Antibody to Canine CD19 for Diagnostic and Future Therapeutic Purposes. Veterinary pathology, 57(2), 241-252.
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