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BigEasy® v2.0 Linear Cloning System (pJAZZ® Vectors)

  • Maximum insert stability.
  • Efficiently clone any insert up to 30 kb.
  • Create libraries from A/T-rich or G/C-rich genomes.
  • Clone gene clusters or operons.
  • Inducible copy number.

The BigEasy Kit with the pJAZZ vector is ideal for constructing bias-free, large-insert genomic libraries, or for cloning difficult DNA of any size up to 30 kb.  Because the novel pJAZZ vector is maintained as a linear molecule, the ends of the vector can rotate freely.  As a result, the vector does not supercoil.  Without the torsional stress induced by supercoiling, otherwise unclonable sequences (e.g., repetitive, or A/T-rich or G/C-rich sequences) are stabilised.  The pJAZZ linear cloning vector is maintained at low copy number (5-10/cell) in BigEasy-TSA™ Electrocompetent Cells (>4 × 1010 cfu/µg), which are required for transformation and propagation. The copy number can be induced 5-10X in this strain. In addition, the pJAZZ vector incorporates Lucigen’s CloneSmart® technology for transcription-free cloning, which further increases insert stability. pJAZZ clones can be isolated using conventional plasmid prep methods and sequenced using standard techniques.

pJAZZ-OC and pJAZZ-OK vectors

Figure 1. pJAZZ-OC and pJAZZ-OK linear vectors. RepA, replication factor and low copy origin of replication (~2-4 per cell; inducible 5-10 fold); Camr - chloramphenicol resistance gene; Kanr - kanamycin resistant gene; telN - protelomerase gene; cB - replication regulator. Approximate positions of transcription terminators (T) are indicated.

Figure 2. Sequence trace of a Piromyces clone, showing extremely high (96%) AT content (see figure 3d).

Figure 3. 15-20 kb PCR amplification products cloned into the pJAZZ® linear vector.

 

Figure 4. Mouse genomic DNA sheared to
6-20 kb and cloned into the pJAZZ linear vector.

 

Figure 5. Oxytricha trifallax genomic DNA (75-85% AT) sheared to 6-20 kb and cloned into the pJAZZ linear vector.

Figure 6. Piromyces genomic DNA (85% AT) cloned in the pJAZZ vector. This DNA was unclonable in all other vectors.

Convenient Success

Efficiently clone the most difficult DNAs

The BigEasy v2.0 Kits eliminate tedious vector and competent cell preparation, as well as time-consuming QC testing.  Kits include optimised reagents, ligation-ready vector, highly efficient electrocompetent cells, detailed instructions, and trouble-shooting guides.

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Product Code Product Name QTY Price Action
60224-2 BigEasy TSA ElectroComp.Cells SOLOS 12rxns
$560.00
60224-3 BigEasy TSA ElectroComp.Cells SOLOS 24rxns
$1,011.00
60224-1 BigEasy TSA ElectroComp.Cells SOLOS 6rxn
$340.00
43018-2 BigEasy v2.0 Linear Cl.Kit OC Blunt 10rxns
$1,106.00
43018-3 BigEasy v2.0 Linear Cl.Kit OC Blunt 20rxns
$1,962.00
43024-2 BigEasy v2.0 Linear Cl.Kit OC Notl 10rxns
$1,106.00
43024-1 BigEasy v2.0 Linear Cl.Kit OC Notl 5rxns
$659.00
43036-2 BigEasy v2.0 Linear Cl.Kit OK Blunt 10rxns
$1,106.00
43036-3 BigEasy v2.0 Linear Cl.Kit OK Blunt 20rxns
$1,962.00
43036-1 BigEasy v2.0 Linear Cl.Kit OK Blunt 5rxns
$659.00
43042-1 BigEasy v2.0 Linear Cl.Kit OK Notl 5rxns
$659.00
43042-2 BigEasy v2.0 Linear Cl.Kit OK Notl 10rxns
$1,106.00
43042-3 BigEasy v2.0 Linear Cl.Kit OK Notl 20rxns
$1,962.00

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