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Fast-Link DNA Ligation Kit

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

Fast-Link DNA Ligation Kit

Ligate DNA for cloning applications with this complete kit.
  • Clone with speed and efficiency
  • Transform into E. coli without clean-up or de-salting
Item ID MBTOOL-010
TBD
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Product information

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 de-salt 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.

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.
Type of End Ligation Time % White Colonies Recombinants per µg DNA
Overhang 5 min. 93 2.0 x 10 6
Blunt 15 min. 71 4.4 x 10 5
PCR product with 3´-A overhang 1 hr. 68 1.2 x 10 4

Table 1. Fast-Link representative results.

Benefits

  • Cohesive-end ligations in 5 minutes at room temperature (Figure. 1).
  • Blunt-end ligations in 15 minutes at room temperature (Figure. 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.

Manuals and user guides

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