What are XL1-Blue competent cells?
XL-1 Blue Competent Cells for routine cloning allow blue-white color screening, single-strand rescue of phagemid DNA, and preparation of high-quality plasmid DNA. Derivatives enable higher transformation efficiency, transformation of methylated DNA, choice of antibiotic resistance, and no F episome.
Are XL1-Blue cells E coli?
Info to: XL1-blue cells (E. coli): XL1-Blue cells are endonuclease (endA) deficient, which greatly improves the quality of Miniprep DNA, and are recombination (recA) deficient, improving insert stability. The hsdR mutation prevents the cleavage of cloned DNA by the EcoK endonuclease system.
How do I make XL1-Blue competent cells?
- Autoclave 100 ml LB- broth in a 1000 ml flask.
- Sterilize and pre-chill the 50-mL conical tubes.
- Prepare 30 ml of ice cold MgCl2-CaCl2 solution (80mM MgCl2, 20 mM CaCl2)
- Prepare ice cold 0.1 CaCl2.
- Pre-warm the LB plates.
How can we preserve competent cells?
Competent cells need to be stored at -80 °C. The process of making the cells competent makes them very fragile – likely to rupture and die. This means that storing at -20 °C can dramatically impede the transformation efficiency.
What is E coli XL1-Blue?
BACKGROUND. The XL1-Blue strain allows blue-white color screening for recombinant plasmids and is an excellent host strain for routine cloning applications using plasmid or lambda vectors.
What is E coli XL1 Blue?
Where do you store XL1 blue competent cells?
XL1-Blue competent cells (blue tubes) 5 × 0.2 ml ≥1 × 108 pUC18 control plasmid (0.1 ng/µl in TE buffer) 10 µl — β-Mercaptoethanol (1.42 M) 25 µl — Storage: Supercompetent cells must be placed immediately at the bottom of a –80°C freezer directly from the dry ice shipping container. Do not store the cells in liquid nitrogen.
How are xl2-blue ultracompetent cells different from XL1-Blue cells?
XL2-Blue ultracompetent cells are high-efficiency derivatives of our XL1-Blue supercompetent cells. XL2-Blue cells are endonuclease deficient (endA), greatly improving the quality of miniprep DNA, and recombination deficient (recA), helping to ensure insert stability.
Why are XL1 blue cells recombinase deficient?
XL-1 cells are tetracycline resistant. XL1-Blue cells are endonuclease (endA) deficient, which greatly improves the quality of miniprep DNA, and are recombination (recA) deficient, improving insert stability. The hsdR mutation prevents the cleavage of cloned DNA by the. EcoK endonuclease system.
Which is the host strain for XL1 blue plasmid?
BACKGROUND The XL1-Blue strain allows blue-white color screening for reco mbinant plasmids and is an excellent host strain for routine clon ing applications using plasmid or lambda vectors. XL1-Blue Genotype: recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F´ proAB lacIqZ∆M15 Tn10 (Tetr)].