The BigEasy Kit with the pJAZZ vector from Lucigen is ideal for constructing bias-free, large-insert genomic libraries, or for cloning difficult DNA of any size up to 30 kb. As the novel pJAZZ vector included is maintained as a linear molecule, the ends of the vector can rotate freely and as a result, the vector does not supercoil. Without the torsional stress induced by supercoiling, previously unclonable sequences such as repetitive and A/T-rich or G/C-rich sequences are stabilised.
Kits include optimised reagents, ligation-ready vector, highly efficent electrocompetent cells, detailed instructions as well as trouble-shooting guides.
Benefits of using the BigEasy v2.0 linear cloning system
• 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
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.
(A) 15-20 kb PCR amplification products cloned into the pJAZZ® linear vector. (B) Mouse genomic DNA sheared to 6-20 kb and cloned into the pJAZZ linear vector. (C) Oxytricha trifallax genomic DNA (75-85% AT) sheared to 6-20 kb and cloned into the pJAZZ linear vector. (D) Piromyces genomic DNA (85% AT) cloned in the pJAZZ vector. This DNA was unclonable in all other vectors.