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BigEasy Linear Cloning Kits (especially advantageous when cloning large DNAs)

Efficiently clone large DNAs up to 30kb
• Clone large inserts from even the toughest DNAs
• 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.

Image
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.

Image
Sequence trace of a Piromyces clone, showing extremely high (96%) AT content.

Image
Oxytricha trifallax genomic DNA (75-85% AT) sheared to 6-20 kb and cloned into the pJAZZ linear vector.
Left arm: 12kb; Right arm: 2kb


BigEasy v2.0 Linear Cloning Kit Contents
- Dephosphorylated pJAZZ vector pre-cut at either a SmaI (blunt) or NotI site
- CloneSmart DNA Ligase
- CloneDirect™ 10X Ligation Buffer (includes ATP)
- Sequencing Primers
- Positive Control Insert DNA
- DNA Terminator® End Repair Enzyme (blunt kits only)
- BigEasy-TSA Electrocompetent Cells in SOLO packaging (1 transformation per tube)
- Recovery Medium
- Transformation Control DNA

For maximum transformation efficiency, use of Lucigen’s BigEasy TSA Electrocompetent Cells is required. BigEasy- TSA Electrocompetent Cells are also available separately, see link below.



Related Links

BigEasy TSA Electrocompetent Cells
PRECISOR High-Fidelity DNA Polymerase


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User Manual BigEasy Linear Cloning Kits

 

 

Description Cat# Size Price    
BigEasy v 2.0 Linear Cloning Kit (pJAZZ-OC Blunt Vector, Cm Resistance) with BigEasy TSA Electrocompetent Cells 43018-1-LU 5 rxns 448 € DETAILS   Add to Cart 
BigEasy v 2.0 Linear Cloning Kit (pJAZZ-OC Blunt Vector, Cm Resistance) with BigEasy TSA Electrocompetent Cells 43018-2-LU 10 rxns 694 € DETAILS   Add to Cart 
BigEasy v 2.0 Linear Cloning Kit (pJAZZ-OC Blunt Vector, Cm Resistance) with BigEasy TSA Electrocompetent Cells 43018-3-LU 20 rxns 1162 € DETAILS   Add to Cart 
BigEasy v 2.0 Linear Cloning Kit (pJAZZ-OC NotI Vector, Cm Resistance) with BigEasy TSA Electrocompetent Cells 43024-1-LU 5 rxns 448 € DETAILS   Add to Cart 
BigEasy v 2.0 Linear Cloning Kit (pJAZZ-OC NotI Vector, Cm Resistance) with BigEasy TSA Electrocompetent Cells 43024-2-LU 10 rxns 694 € DETAILS   Add to Cart 
BigEasy v 2.0 Linear Cloning Kit (pJAZZ-OK Blunt Vector, Kan Resistance) with BigEasy TSA Electrocompetent Cells 43036-1-LU 5 rxns 448 € DETAILS   Add to Cart 
BigEasy v 2.0 Linear Cloning Kit (pJAZZ-OK Blunt Vector, Kan Resistance) with BigEasy TSA Electrocompetent Cells 43036-2-LU 10 rxns 694 € DETAILS   Add to Cart 
BigEasy v 2.0 Linear Cloning Kit (pJAZZ-OK Blunt Vector, Kan Resistance) with BigEasy TSA Electrocompetent Cells 43036-3-LU 20 rxns 1162 € DETAILS   Add to Cart 
BigEasy v 2.0 Linear Cloning Kit (pJAZZ-OK NotI Vector, Kan Resistance) with BigEasy TSA Electrocompetent Cells 43042-1-LU 5 rxns 448 € DETAILS   Add to Cart 
BigEasy v 2.0 Linear Cloning Kit (pJAZZ-OK NotI Vector, Kan Resistance) with BigEasy TSA Electrocompetent Cells 43042-2-LU 10 rxns 694 € DETAILS   Add to Cart 
BigEasy v 2.0 Linear Cloning Kit (pJAZZ-OK NotI Vector, Kan Resistance) with BigEasy TSA Electrocompetent Cells 43042-3-LU 20 rxns 1162 € DETAILS   Add to Cart 

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