Standard cloning for large inserts and T1 phage resistance

• Avoid phage contamination with tonA mutation for resistance to bacteriophages T1 and T5.
• Clone large inserts and transform large plasmids - up to at least 145 kb plasmid DNA.
• Generate clones with inducible copy number using CopyControl™ vectors.
• Achieve high transformation efficiencies: >1 × 10¹⁰ cfu/µg pUC19 DNA.

Applications

• Construction of inducible-copy-number genomic libraries using the CopyControl™ Cloning System, with clones that are resistant to contaminating phage T1 and T5.

Phage T1-Resistant TransforMax™ EPI300™-T1^RE. coli have all the benefits of the highly versatile TransforMax EPI300™ E. coli competent cells with the addition of being resistant to bacteriophages T1 and T5 (tonA genotype). Once introduced into the lab environment, bacteriophage T1 rapidly lyses E. coli strains that are commonly used in cloning applications and results in significant lab downtime and the loss of valuable clones. Bacteriophage T1 is particularly difficult to eliminate from the lab and can lay dormant for many years. The tonA genotype protects the phage T1-resistant TransforMax EC100-T1^R cells and valuable clones from attack by bacteriophage T1.

Like the standard TransforMax EPI300 E. coli, this strain has been specially engineered for use with Epicentre's CopyControl™ Cloning Systems.* The cells contain an inducible mutant trfA gene whose gene product is required for initiation of replication from the oriV contained on the CopyControl pCC1™ Vectors or on clones that have been retrofitted with CopyControl capability using the EZ-Tn5™<oriV/KAN-2> Insertion Kit. This process allows TransforMax EPI300-T1^RE. coli to maintain CopyControl vectors at single copy until induction occurs immediately before DNA purification.

Benefits

• tonA for resistance to bacteriophages T1 and T5.
• trfA gene under tight control of an inducible promoter for copy-number control of CopyControl clones and clones retrofitted with the EZ-Tn5<oriV/KAN-2> Transposon.
• High transformation efficiency of both large and small clones.
• lacZΔM15 for blue/white screening of recombinants.
• Readily accepts large DNAs for construction of large-insert genomic libraries.
• Restriction minus [mcrA, Δ(mrr-hsdRMS-mcrBC)] enables efficient cloning of methylated DNA.
• Endonuclease minus (endA1) to ensure high yields of DNA.
• Recombination minus (recA1) for greater stability of large cloned inserts.

Genotype

F^- mcrA Δ(mrr-hsdRMS-mcrBC) Φ80dlacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara, leu)7697 galU galK λ^- rpsL (Str^R) nupG trfA tonA

*Covered by issued and/or pending patents.