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BigEasy® v2.0 Linear Cloning Kit (pJAZZ®-OK Blunt Vector)
BigEasy® v2.0 Linear Cloning Kit (pJAZZ®-OK Blunt Vector)
货号
43036-3
规格
20 rxns
存储温度
-20º & -80º C
产品详情
- 最大的插入稳定性
- 有效克隆任意插入30kb片段
- 以A/T或G/C含量丰富的基因组创建文库
- 克隆基因簇或操纵子
- 诱导拷贝数
The BigEasy Kit 带有pJAZZ载体，是理想化构建无偏好性，较大插入基因组文库或者用于克隆30kb的困难DNA。因为新的pJAZZ载体可作为线型分子来存在，载体末端可以自由旋转。但载体不能超螺旋。在没有超螺旋扭曲压力诱导下，序列可以保持稳定性，尤其是重复序列或者AT和GC含量丰富的序列。pJAZZ线性克隆载体在BigEasy-TSA™ 电转化感受态细胞 (>4 × 1010 cfu/µg) 中是低拷贝的，约每个细胞5-10个拷贝，利于转化和增殖。另外，pJAZZ载体整合了Lucigen’s CloneSmart®技术用于无转录克隆，增加插入稳定性。 pJAZZ克隆可以使用传统的质粒提取方法来提取，并使用标准技术进行测序。见pJAZZ高通量测序手册。
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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. |
Convenient Success
有效克隆困难DNA
The BigEasy v2.0 Kits 消除了繁琐的载体和感受态细胞的制备，也省去了QC试验所耗费的时间。盒子包括最优化的试剂，连接载体，高效电转感受态细胞，详细的操作说明书和疑惑解答。
参考文献
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 >>
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Ordering
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Manuals
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Resources
BigEasy v2.0 Linear Cloning Kits 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.
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