TFB Electro 10B™ in Cuvettes
TFB Electro 10B™ in Cuvettes™ are pre-dispensed in sterile, 1 mm gap electroporation cuvettes for simple, convenient electro-transformation. The high transformation efficiency of these cells makes them ideally suited for demanding applications such as cDNA or genomic library construction.
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| Recommended Use | cDNA or genomic library construction, generation of ssDNA, cloning and expression of toxic genes, cloning of methylated DNA. |
| Efficiency | ≥ 10<sup>10</sup> cfu / µg |
| Shipping | Dry ice |
| Genotype | F<sup>-</sup> <i>mcr</i>A Δ(<i>mrr-hsd</i>RMS-<i>mcr</i>BC) Φ80/acZΔM15 Δ<i>lac</i>X74 <i>rec</i>A1 <i>end</i>A1 <i>ara</i>D139 Δ(<i>ara, leu</i>)7697 <i>gal</i>U <i>gal</i>K <i>rps</i>L <i>nup</i>G λ<sup>-</sup> <i>fhu</i>A (T1<sup>R</sup>). Note: The Δ(<i>mrr-hsd</i>RMS-<i>mcr</i>BC) deletion, together with the mcrA mutation, allow cloning of methylated DNA such as genomic DNA from eukaryotic sources. |
Description
TFB Electro 10B™ in Cuvettes™ are pre-dispensed in sterile, 1 mm gap electroporation cuvettes for simple, convenient electro-transformation. The high transformation efficiency of these cells makes them ideally suited for demanding applications such as cDNA or genomic library construction (1). TFB Electro 10B™ 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 -70o or below.
- Competent cells should be thawed on wet ice (~ 10 minutes) and should be used promptly after thawing.
- Thawed cells can be refrozen by placing them in dry ice although a drop in transformation efficiency of as much as ≥ 50% may be observed.
- Cells stored properly will remain stable for up to 2 years.
Genotype
F- mcrA Δ(mrr-hsdRMS-mcrBC) Φ80/acZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara, leu)7697 galU galK rpsL nupG λ- fhuA (T1R).
Note: The Δ(mrr-hsdRMS-mcrBC) deletion, together with the mcrA mutation, allow cloning of methylated DNA such as genomic DNA from eukaryotic sources.
Quality Control
Each lot of TFB Electro 10B™ cells is tested to verify transformation efficiencies of ≥1 x 1010 transformants/ug with limiting amounts of supercoiled pUC19 DNA (10 pg). Saturating amounts of DNA (250ng) give ≥1 x 108 colonies/ 20µl reaction. 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:
