LiClone™ Fast Cloning Kits

LiCloneTM One Step DNA Assembly Kit is a novel cloning kit, applicable for homologous recombination with 1 - 5 fragments. The Kit is independent of DNA ligase, significantly reducing the self-ligated colonies. The enhanced Exnase and highly optimized buffer included in 2× LiClone Mix significantly improve the recombination efficiency and the tolerance to impurities. pCE-Zero vector is compatible with most PCR products, enabling the specific PCR products to be used directly for recombination without any treatments, which significantly simplify the procedure.

Key Features

• Simple: one-tube super mix.
• Fast: cloning within 5 - 15 min.
• Efficient cloning of fragments of 50 bp-10 kb, with positive clone rate > 95%.
• Directional cloning at any site on any vector, no limits in restriction enzyme choice.
• Suitable for cloning of both single and multiple DNA fragments.

Applications

• Fast Cloning
• High-throughput Cloning
• Seamless Assembly
• Site-directed Mutagenesis

Principal

Acquisition of Inserts: Introduce URS/DRS sequences (highlighted in blue and red ) into 5’-end of Forward (F) & Reverse (R) primers, aiming to make the ends of amplified inserts and pCE-Zero vector identical to each other. (Step I). Recombination: Mix pCE-Zero vectors and inserts at an appropriate ratio and incubate with Exnase at 50°C for 5min to make two linearized DNA cyclized in vitro. (Step II). Transformation: The recombination products can be used for transformation directly(Step III).

Customer Testimonials

"Our lab had a chance to try the LiTaq™ Plus PCR Master Mix and LiClone™ One Step DNA Assembly Kit which perform very well compared with similar products we used before. We will order them later. Thank you!"

Product Name	Cat. #	Size	Price	Order
LiCloneTM One Step DNA Assembly Kit	M0010-25	25 rxn	$199  $159
LiCloneTM Fast DNA Ligation Kit	M0009	100 rxn	$199

Case Study

LiClone™ One Step DNA Assembly Kit can clone 50bp-10kb fragment, single fragment with different length (50bp, 2kb, 4.5kb, 7.5kb, 10kb), multiple fragments: four fragments (3kb), three fragments (10kb), five fragments (3kb) and five fragments (6kb) with high efficiency; after purification can be homologously recombinated with pCE-Zero vector; recombination products will be used to transform competent cell DH5α. This kit is independent with ligase, self-link without vector and the positive cloning rate can be as high as 95% or above. After transformation, 50 clones are identified by PCR, the positive rates of insertion with different fragment length are shown right.

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