BigEasy Linear Cloning Kits
BigEasy Linear Cloning Kits - especially advantageous when cloning large DNAs
Efficiently clone DNAs up to 30kb
- Benefit from transcription- and translation-free cloning
- Achieve highest insert stability due to linear pJAZZ vectors
- Prevent insert size or content bias
The BigEasy v2.0 Linear Cloning Kits are based on a novel linear cloning vector (patents pending) derived from the linear phage N15 of E. coli. Due to its unique replication system, the pJAZZ vector behaves like a linear plasmid in BigEasy-TSA™ Electrocompetent Cells. Because the pJAZZ vector is linear, the ends of the plasmid can rotate freely. Therefore, it is not under torsional stress and inserts are much more stable, enbaling the successful cloning even of AT-rich sequences or inverted repeats. Added insert stability is achieved by the presence of CloneSmart transcription terminators and a promoter-free insertion site in the pJAZZ vectors, which prevent transcription/translation of toxic or deleterious sequences. Thus, large inserts from even the toughest DNAs, including those with extensive nucleotide repeats and extremely high AT content, can be successfully cloned.
The BigEasy v2.0 Linear Cloning Kits are ideal for constructing bias-free, large insert genomic libraries, or for cloning difficult DNA of any size up to 30 kb. They are particularly useful when the target DNA may contain regions that are unclonable in conventional BAC or other vectors, including high AT content, toxic genes, strong promoters, or nucleotide repeats.
Fig.1. Schematic diagram of the 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.
The pJAZZ vectors contain a low copy, inducible origin of replication. The copy number is ~2-4/cell prior to induction; it is increased by approximately 5-10 fold in BigEasy-TSA cells by addition of the Induction Solution provided in the BigEasy Kit.