BigEasy® v2.0 Linear Cloning System (pJAZZ® Vectors): pJAZZ-OC
- Efficiently clone any insert up to 30 kb.
- Create libraries from even A/T- or G/C-rich genomes.
- Clone gene clusters or operons.
- Maximum insert stability.
- Inducible copy number.
Ordering
BigEasy v2.0 Linear Cloning Kit include: 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, BigEasy-TSA Electrocompetent Cells in SOLO packaging (1 transformation per tube), Recovery Medium, Transformation Control DNA, and complete protocols. BigEasy-TSA Electrocompetent Cells are also available separately.
Description
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 linear, 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- 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 x 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. The pJAZZ vector can be isolated using standard plasmid prep methods.
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Figure 1. The pJAZZ-OC and pJAZZ-OK linear vectors. RepA - replication factor and low copy origin of replication (~5-10 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. |
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Figure 2. Sequence trace of a Piromyces clone from Figure 6, showing extremely high (96%) AT content. |
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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 obtain your clone or create your library
The BigEasy v2.0 Kits eliminate tedious vector and competent cell preparation, as well as time-consuming QC testing experiments. Kits include optimized reagents, ligation-ready vector, highly efficient electrocompetent cells (not applicable to fosmid kits), detailed instructions, and trouble-shooting guides to simplify cloning and sequencing.
Resources
| Document | File Name | Type |
| Manual | MA033-BigEasy-v20-Linear-Cloning-Kit | |
| Manual | MA042-BigEasy-TSA-Electrocompetent-Cells | |
| Vector Sequence | Sequence(s) | Link |
| MSDS |
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