Cloning kit with higher-efficiency competent cellsThe pSTBlue-1 Perfectly Blunt® Giga Cloning Kits feature NovaBlue GigaSingles™ Competent Cells for increased transformation efficiency (> 1 × 109 cfu/µg). NovaBlue is a K-12 strain ideally suited as an initial cloning host due to its high transformation efficiency, blue-white screening capability (with appropriate plasmids) and recA endA mutations, which facilitate high yields of excellent-quality plasmid DNA. Cells are provided in a 50-µl, single-use volume that eliminates the need to dispense, freeze/thaw or waste partially used vials.The Perfectly Blunt Cloning Kits are designed for simplified cloning of DNA generated by PCR using any type of DNA polymerase. This approach enables the use of high-fidelity proofreading enzymes for amplification, thus decreasing the probability of generating mutations in the target sequence. In addition, under many conditions blunt cloning is more efficient than T-cloning, most likely due to the observation that the efficiency of single dA addition by Taq DNA polymerase varies significantly depending on the sequence context of the DNA ends, and even the number of PCR cycles performed (1–5).Note that the Perfectly Blunt method is not limited to cloning PCR products; these kits are also suitable for cloning restriction fragments, cDNA, or sheared DNA with the same protocols.The pSTBlue-1 vector is available in a kit containing sufficient reagents for 20 or 40 reactions. "Vector only" kits are also available in 20 and 40 reaction sizes without ligase and competent cells.
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Product information
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Components
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| • | 2 or 4 × 0.5 µg | pSTBlue-1 Blunt Vector
| | • | 45 ng | Blunt Vector Positive Control Insert
| | • | 1 or 2 × 100 U | T4 DNA Ligase
| | • | 1 or 2 × 0.1 ml | End Conversion Mix
| | • | 1.5 ml | Nuclease-free Water
| | • | 22 or 44 × 50 µl | NovaBlue GigaSingles™ Competent Cells
| | • | 4 or 7 × 2 ml | SOC Medium
| | • | 10 µl | Test Plasmid |
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Features and benefits
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- No restriction enzymes or special primers
- Compatible with any DNA polymerase
- Independent of 3′-dA addition or sequence context of DNA ends
- Blue/white screening with seven different vectors, including pETBlue expression vectors
- Simple protocol takes less than one hour* from PCR product to plating transformants
* All times listed refer to kits using NovaBlue Singles™ Competent Cells and ampicillin or chloramphenicol selection. Giga kits, which use NovaBlue GigaSingles™ Competent Cells, require one hour for outgrowth.
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Store and ship information
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Storage
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≤ -70°C
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Ship
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Dry Ice Only
Multiple Toxicity Values, refer to MSDS
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