pSpark® Done

For highly Efficient, Accurate & Easy Cloning of PCR fragments with EcoRI and NotI flanking the insertion site

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.

Price: 196.54 

SKU: C0006 Categories: ,

Detailed information:

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.

– 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)

Download documentation
  • 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.
Tables & Figures


Quality Control
  • Functional test using a 1.0 kb PCR fragment.
  • Recommendations: All pSpark® DNA cloning vectors are stable for at least 1 month at 4 °C and even at 20 – 25 °C for up to 2 days although storage temperatures above -20 °C are not recommended. In case of incident, such as a power failure, stored vectors should be tested with the supplied control insert before considering discarding it. However, please note that T4 DNA Ligase is extremely temperature-sensitive and storage temperatures above    -20 °C inactivates the enzyme.
Storage, Shipping & Guarantee
  • Shipped in: Gel Pack.
  • Storage: -20 ºC (NON Frost-Free Freezer).
Safety Statements

This product is developed, designed and sold exclusively for Research purposes and in vitro use only (RUO). The product was not tested for use in diagnostics or for drug development, nor is it suitable for administration to humans or animals. For more info, please check its Material Safety Data Sheet available in this website.

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