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BigEasy® v2.0 Linear Cloning Kit (pJAZZ®-OC Blunt Vector)
大量
中北林格
5 rxns
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. See protocol for high-throughput Sanger sequencing of pJAZZ clones.
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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. |
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Figure 2. Sequence trace of a Piromyces clone, showing extremely high (96%) AT content (see figure 3d). |
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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. |
The BigEasy v2.0 Kits eliminate tedious vector and competent cell preparation, as well as time-consuming QC testing. Kits include optimized reagents, ligation-ready vector, highly efficient electrocompetent cells, detailed instructions, and trouble-shooting guides.
Published Paper:
Godiska R, Mead DA, Dhodda V, Hochstein R, Karsi A, Usdin K, Entezam A, Ravin N. (2010) Linear plasmid for cloning large or repetitive sequences in E. coli. Nuc. Acids Res. 38:e88. Read Paper >>
Product Description | Size | Cat. No. | |
BigEasy v2.0 Linear Cloning Kit (pJAZZ-OC NotI Vector) | 5 rxns | 43024-1 | |
10 rxns | 43024-2 | ||
20 rxns | 43024-3 | ||
BigEasy v2.0 Linear Cloning Kit (pJAZZ-OK Blunt Vector) | 5 rxns | 43036-1 | |
10 rxns | 43036-2 | ||
20 rxns | 43036-3 | ||
BigEasy v2.0 Linear Cloning Kit (pJAZZ-OK NotI Vector) | 5 rxns | 43042-1 | |
10 rxns | 43042-2 | ||
20 rxns | 43042-3 | ||
BigEasy® v2.0 Linear Cloning Kit (pJAZZ-OC Blunt Vector) | 5 rxns | 43018-1 | |
10 rxns | 43018-2 | ||
20 rxns | 43018-3 | ||
BigEasy-TSA Electrocompetent Cells | 6 rxns (SOLOs) | 60224-1 | |
12 rxns (SOLOs) | 60224-2 | ||
24 rxns (SOLOs) | 60224-3 |
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