pSpark® II

pSpark® II is a highly efficient, accurate and easy-to-use DNA Cloning system based on a breakthrough technology for cloning blunt ended DNA generated by PCR with a proofreading or high fidelity DNA polymerases.

The vector is prepared by digestion of pSpark® II at the EcoRV site before treating both ends to prevent vector self-ligation. The end treatment is supported by a proprietary know-how that guarantees a higher cloning efficiency than with simply a desphorylated vector.

Advantages & Features
- Unprecedented high cloning efficiency: more than 2,500 positive colonies expected under optimal conditions.
- Great efficiency: over hundreds positive colonies with few nanograms of insert.
- High stability: eliminates cloning bias or pitfalls.
- Time-saving protocol: no hidden steps such as phosphorylation, just ligation after PCR and transformation.
- Powerful: clone from less than 1 ng/kb to up to 14 kb; obtain 4-fold more positive colonies using 3-fold less DNA insert.
- Easy-to-use: eliminate recombinant screening due to its minumum background (lower than 1%), avoiding “suicide” strategies from toxic genes.
- Great versatility: compatible with any protocol, proofreading Polymerase, Competent Cells, ligation time or primers.
- Flexible: ligation time from 10 minutes to overnight.
- Robust for every DNA size: just 6.7 ng per kb of insert needed for optimal ligation.
- High cost-saving: reduces your cloning costs as no expensive phosphorylated primers are needed.
- Eliminates positive selection vector.

Includes
- 20 rxn pSpark® II (20 ng/µL)
- 20 µL T4 DNA Ligase (5 Weiss U/µL)
- 100 µL T4 DNA Ligase Buffer (10x)
- 150 µL PEG 6000 (10x)
- 5 µL Insert Control 1 kb (20 ng/µL)

Applications
- General cloning.
- Clone PCR fragments included in a low amount.
- Cloning of PCR products amplified with High Fidelity polymerases.
- Cloning of PCR fragments generated with a blend of polymerases.
- Production of ssDNA.
- Blue/white screening for recombinants.
- In vitro transcription from T7/SP6 dual-opposed promoters.

Shipping & Storage
Shipped in: Gel pack.
Storage: -20 ºC (NON Frost-Free Freezer).