Intact Genomics (IG^®) propriety Quick10^™ Cloning kit and DirectPlate^® technologies enable simple, rapid and highly efficient cloning. The Quick10^™ Cloning kit utilizes homologous assembly which allows for nearly 100% cloning accuracy. This kit enables the direct cloning of one or two DNA fragment(s) (<6kb total) through PCR into linearized cloning-ready Ig^® AmpR/KanR-pCR vector (4 kb, provided in this kit). The PCR fragments can be generated by Intact Genomics high fidelity i7^® DNA polymerase or other high fidelity DNA polymerases. Primers need to have 12 to 15 bases of homology at Ig^® AmpR/KanR-pCR vector linear ends where the DNA fragment(s) or PCR product(s) needs to fuse. The unpurified PCR product (s) is then diluted 5-10 times by the dilution buffer included in this kit. No purification step is required. The diluted PCR product (s) and linearized Ig^® AmpR/KanR-pCR vector can then be assembled in just 7 minutes at 37 ºC, followed by a 3 minutes transformation step on ice utilizing our Directplate^® XL DH10B competent cells (included in this kit).  DirectPlate^® cells eliminate heat shock, lengthy incubations, and time-consuming outgrowth procedures. Utilizing this kit, DNA/PCR product(s) can be assembled/transformed in 10 minutes vs. other seamless/fusion cloning, conventional T/A, or restriction ligation cloning and transformation methods that can take up to 2 hours or more.

Key Benefits

• Combination of high fidelity PCR, assembly, and DirectPlate^® transformation cloning technologies.
• Combination of homologous assembly and ccdB selection (vs. T/A or restriction ligation methods) displays <1% false-positive clones or nearly 100% cloning accuracy.
• Assembled/transformed in 10 minutes. Less time and less effort spent on cloning, transformation, and positive clone screening/identification.
• Single selection vector or custom vector available upon request

Applications

• Streamlined cloning of one or two (<6 kb total) DNA fragments
• Single PCR product cloning
• Site directed mutagenesis
• High throughput cloning

Product Includes1)  5x Quick10-Assemble^™ Premix2)  Ig^® AmpR/KanR-pCR vector – cloning ready3)  Dilution buffer4)  Directplate^® XL DH10B competent cells (6×50 µl)

Storage Temperature-20 ºC for premix, vector and buffer-80 ºC for Directplate^® XL DH10B competent cells

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Please find the video below describing full use of this kit.

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