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 stabilized. 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.
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.
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Figure 4. Mouse genomic DNA sheared to
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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.