TFB 5 Alpha™ - 96-Well

TFB 5 Alpha™ (TFB5α™) Competent Cells are chemically competent E. coli cells and are intended to be transformed by heat-shock. The high transformation efficiency and 96-well formatting of these cells make them ideally suited for high-throughput cloning applications.  TFB5α™ cells are resistant to bacteriophage T1, an especially virulent virus which attacks E. coli, and can be rapidly spread via aerosols.

Handling and Storage

• Upon receipt, store cells immediately at -70°or below.
• Competent cells in 96-well plates should be thawed in a pre-chilled (0°C) metal heating block or on wet ice (~ 5 minutes) and used promptly after thawing.
• Cells stored properly will remain stable for up to 2 years.

Kit Contents

• TFB 5α™ Competent Cells
• pUC19 Control DNA (10 pg / µl)

Genotype

F^-Φ80dlacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17(r^-_k, m⁺_k) phoA supE44 λ^- thi-1 gyrA96 relA1 fhuA (T1^R).

Quality Control

TFB 5α™ cells are tested to verify transformation efficiencies of ≥ 1 x 10⁸ transformants/µg with limiting amounts of supercoiled pUC19 DNA (20 pg). Untransformed cells are also tested for resistance to bacteriophage T5, a standard test for resistance to phage T1, as well as other relevant genetic markers and sensitivities to antibiotics.

Considerations in Choosing Competent Cells

Key Genetic Markers:

• recA - eliminates homologous recombination; stabilizes inserts.
• endA - endonuclease deficiency improves yield and quality of plasmid DNA.
• hsd - classical restriction/modification system, deficiency allows cloning from non-E. coli sources, e.g. PCR products.
• F’ - episome required for infection by M13 and other phagemids. Important in generation of ssDNA.
• mcrA, mcrBC, mrr - mutations in these genes allows efficient cloning of genomic from mammalian or other higher eucaryotes, and methylated cDNA.
• lacZΔM15 - omega donor; allows a-complementation for blue/white screening.

Product Selection Guide:

• Competent Cells Selection Guide