pSpark® Done

pSpark® Done is a highly efficient, accurate and easy-to-use DNA Cloning system designed for cloning of blunt ended DNA with very high efficiency. The MCS of the pSpark® Done vector incorporates sequences on either side of the insert that are recognized by the restriction enzymes NotI and EcoRI. This allows the insert DNA to be removed with a single restriction digest using either of these enzymes.

Advantages & Features
- Optimized: recognition sites for Not I and EcoR I at either side for the insertion of the cloning point.
- Flexible: allows removing the desired insert DNA with other restriction digestion.
- Unprecedented efficiency: more than 2,500 positive colonies expected under optimal conditions.
- Easy-to-use: eliminate screening of recombinants due to its its minumum background (lower than 1%).
- Time-saving protocol: avoids any step required after PCR, just 19 minutes from PCR to plating.
- Powerful: obtain 5-fold more positive colonies using 10-fold less DNA insert.
- High stability: eliminates cloning bias or pitfalls.
- Great versatility: compatible with any protocol, proofreading polymerase, competent cells, ligation time or primers.
- Sensitive: clone from 50 bp insert to up to 14 kb with just 5 ng per kb of insert.
- Eliminates positive selection vector.
- Cost avoidance: removes expensive primer phosphorylation use.
- Robust for every DNA size: just 6.7 ng per kb of insert needed for optimal ligation.

Includes
- 20 rxn pSpark® Done (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
- Cloning of High Fidelity PCR amplified products.
- Production of ssDNA.
- Blue/white screening for recombinants.
- In vitro transcription from T7/SP6 dual-opposed promoters.
- One restriction enzyme allows gene fragment excision.

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