Cambio - Excellence in Molecular Biology

Competent Cells

Competent Cells: Competent cells

A range of electrocompetent and chemically competent cleverly designed E. coli strains for everyday cloning needs through to more demanding applications like CRISPR, working with unstable DNA or low copy number clones. 



TransforMax™ EPI300™ Electrocompetent or Chemically Competent E. coli

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. Increase Gibson Assembly® colony counts with high-efficiency electroporatio

BioSearch Tech (Lucigen/Epicentre)

Description

Insert Priming Sites and Convert Single-Copy Clones to Multi-Copy-Inducible 

The EZ-Tn5™ <oriV/KAN-2> Insertion Kit is designed to simplify and speed up both DNA purification and sequencing of existing single-copy BAC and fosmid clones by integrating CopyControl™ capability. Use a short, one-step in vitro reaction to randomly insert a single EZ-Tn5™ <oriV/KAN-2> Transposon containing an inducible, high-copy origin of replication (oriV) into the target DNA. Then, transform TransforMax™ EPI300™ or EPI300v-T1R Electrocompetent E. coli (sold separately) with an aliquot of the reaction and select for the kanamycin marker encoded by the EZ-Tn5™ Transposon. 

One reaction will generate thousands of EZ-Tn5 <oriV/KAN-2> Transposon insertion clones. Each clone not only contains oriV and a kanamycin marker, but primer binding sites at the ends of the inserted transposon for sequencing. Amplify selected clones by adding the CopyControl™ Induction Solution and purify microgram quantities of template DNA from as little as 1ml of culture. Use the primers provided in the kit to sequence bidirectionally from the ends of the transposon and save the time and money usually spent subcloning or making custom primers.

Figure 1. The process for generating EZ-Tn5 oriV/KAN-2 Transposon insertion clones for high yields of DNA and bidirectional sequencing. Figure 1. The process for generating EZ-Tn5™ <oriV/KAN-2> Transposon insertion clones for high yields of DNA and bidirectional sequencing.



If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Protocols

Protocols for: EZ-Tn5™ <oriV/KAN-2> Kits 

MA172E-TMax-EPI300-cellsl

Please note: all protocols off site are the responsibility of the products supplier

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Notes

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If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Applications & Benefits

Applications

  • Insertion of priming sites and conversion of single-copy clones to multi-copy-inducible clones for higher yields of BAC or fosmid DNA.

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200