• Clone with speed and efficiency
• Transform into E. coli without clean-up or desalting

Applications

• Blunt-end and TA cloning of PCR products.
• Ligation of next-gen sequencing adaptors to blunt-ended DNA.
• Genomic DNA cloning and subcloning.
• BAC/fosmid library construction.
• cDNA cloning.
• Linker ligation.

The Fast-Link™ DNA Ligation Kit uses a high-quality ligase (Fast-Link T4 DNA Ligase), that was cloned at Epicentre and formulated to provide extremely rapid, high-efficiency DNA ligation. Cohesive-end ligations can be performed in 5 minutes at room temperature. In contrast to other ligases, it is not necessary to desalt Fast-Link ligation reactions prior to transformation of electrocompetent or chemically competent cells. The Fast-Link Kit can be used for routine and high-throughput DNA cloning.

Type of End Ligation Time % White Colonies Recombinants per µg DNA Overhang 5 min. 93 2.0 x 106 Blunt 15 min. 71 4.4 x 105 PCR product with 3´-A overhang 1 hr. 68 1.2 x 104 Table 1. Fast-Link™ representative results.

Benefits

• Cohesive-end ligations in 5 minutes at room temperature (Fig. 1).
• Blunt-end ligations in 15 minutes at room temperature (Fig. 2).
• Ligation of PCR product with A-overhangs in 1 hour at 16°C.
• High ligation efficiency (Table 1).
• Desalting of ligation products is not needed prior to transformation of electrocompetent or chemically competent cells.
• BAC ligation with inserts >100 kb can be performed in 4 hours, instead of overnight reactions as with other ligases.

Figure 1. Time course for cohesive end ligation using the Fast-Link™ Kit. Lambda Hind III markers were ligated using 2 units of Fast-Link DNA Ligase (Lanes 2-6). Lane M, Kilobase ladder; lane 1, no enzyme.

Figure 2. Time course for blunt-end ligation using the Fast-Link™ Kit. pUC 19 DNA digested with Pvu II was ligated using 2 units of Fast-Link DNA Ligase (lanes 1-6). Lanes M, Kilobase ladder.

ORDER INFORMATION

Contents: Fast-Link™ DNA Ligase, Fast-Link™ 10X Ligation Buffer, 10 mM ATP.

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