What are XL1 Blue competent cells?

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.

How do I make XL1 Blue competent cells?

1. Autoclave 100 ml LB- broth in a 1000 ml flask.
2. Sterilize and pre-chill the 50-mL conical tubes.
3. Prepare 30 ml of ice cold MgCl2-CaCl2 solution (80mM MgCl2, 20 mM CaCl2)
4. Prepare ice cold 0.1 CaCl2.
5. Pre-warm the LB plates.

How do you resuspend competent cells?

Allow the resuspended cells to chill on ice for ~ 15 minutes. Collect the washed cells by centrifugation for 10 minutes at ~ 7000 rpm at 4 °C and discard the supernatant. Add 5 mL of sterile, ice-cold 100 mM CaCl2 supplemented with 15 % v/v glycerol to each cell pellet and gently resuspend them.

How can we preserve competent cells?

Competent cells should be stored at -80°C. Storage at -20°C will result in a significant decrease in transformation efficiency (TE). When tested on NEB 5-alpha Competent E. coli (NEB #C2987H), cells lost 94.5% of TE after only 24 hours of storage at -20°C.

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.

Are XL1 Blue cells E coli?

What are Electrocompetent cells?

Electrocompetent cells are made competent using an electrical pulse from an electroporator to create temporary pores (poration) in the cell membrane of either prokaryotic or eukaryotic cells.

What is chemically competent cells?

Chemically competent cells are calcium chloride treated to facilitate attachment of the plasmid DNA to the competent cell membrane. The competent cell is alternatively heated in a water bath, this opens the pores of the cell membrane allowing entry of the plasmid.

Can I store competent cells at?

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.

How long can you store competent cells?

How Long can Competent Cells be Stored. When stored and handled properly, GoldBio competent cells should be stable at -80°C for at least 1 year.

What is the volume of xlxl1-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.

Why choose the right competent cells?

The right competent cells are critical to the success of your workflows, be it general protein expression or cloning, difficult cloning, or expressing toxic proteins. Find the best option for your applications from our extensive and innovative selection of competent cells, electrocompetent or chemically competent cells.

Is XL1-Blue a good host strain?

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)].