- Description 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 fragments (<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 fragments or PCR products need to fuse. The unpurified PCR products are then diluted 5-10 times by the dilution buffer included in this kit. No purification step is required. The diluted PCR products and linearized IG® AmpR/KanR-pCR vector can then be assembled in just 7 minutes at 37 ºC, followed by a 3-minute 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 products 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 Includes 1) 5x Quick10-Assemble™ Premix 2) Ig® AmpR/KanR-pCR vector – cloning ready 3) Dilution buffer 4) 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 4122 Please find the video below describing full use of this kit. 4122
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