pSpark® III
For highly Efficient, Accurate & Easy Cloning with Ampicillin and Kanamycin resistance
pSpark® III is a highly efficient, accurate and easy-to-use DNA Cloning System that combines Ampicillin and Kanamycin resistance. Ideal for cloning PCR products amplified from any plasmid vector without the need to gel-purify bands to eliminate the background due to the template vector used for PCR.
Price: 175.95 €
Detailed information:
Advantages & Features
- Unprecedented high cloning efficiency: more than 2,500 positive colonies expected under optimal conditions.
- Time-saving protocol: no hidden steps such as phosphorylation, just ligation after PCR and transformation.
- Powerful: obtain 5-fold more positive colonies using 10x less DNA insert.
- Easy-to-use: eliminate recombinant screening due to its minumum background (lower than 1%), avoiding “suicide” strategies from toxic genes.
- 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.
- High cost-saving: reduces your cloning costs as no expensive phosphorylated primers are needed.
- Eliminates positive selection vector.
Specifications
Includes
– 20 rxn pSpark® III (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 directly from PCR using plasmid cloned genes as template.
- Unpurified PCR cloning.
- Cloning of high fidelity PCR amplified products.
- Production of ssDNA.
- Blue/white screening for recombinants.
- In vitro transcription from T7/SP6 dual-opposed promoters.
Tables & Figures
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Quality Control
- Functional test using a 1.0 kb PCR fragment.
Advice
- 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).
Citations
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- Bravo Ruiz, G. A. (2013). Sistemas hidrolíticos de componentes vegetales en el patógeno de tomate Fusarium oxysporum f. sp. lycopersici: lipasas y poligalaturonasas.
- Chamizo-Ampudia, A., Galvan, A., Fernandez, E., & Llamas, A. (2011). The Chlamydomonas reinhardtii molybdenum cofactor enzyme crARC has a Zn-dependent activity and protein partners similar to those of its human homologue. Eukaryotic cell, 10(10), 1270-1282.
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.