Diagenode Tagmentase (Tn5 transposase) - unloaded

Diagenode Tagmentase is a hyperactive Tn5 transposase with the potential to enhance epigenetic studies. Its ability to cut DNA and insert sequences of interest in one step makes it the perfect companion for Next-Generation Sequencing experiments using powerful technologies such as ATAC-seq, ChIPmentation, CHANGE-seq and other. The enzyme is not loaded with DNA oligos, providing flexibility of application. To ensure optimal results the concentration may be adjusted with Diagenode Tagmentase Dilution Buffer (Cat. No. C01070011), available separately.

Looking for loaded Tagmentase? Please go to Tagmentase (Tn5 transposase) - loaded.

 Quality control

Figure 1: Efficient fragmentation of the lambda DNA after incubation with the Tagmentase

For fragmentation, 100 ng of DNA from bacteriophage lambda were incubated with diluted Diagenode Tagmentase (Cat. No. C01070010) loaded with sequencing adapters and Tagmentation buffer (1x) (Cat. No. C01019042) for 7 min at 55°C. The reaction was stopped by addition of SDS (0.2% final concentration). After clean-up using AMPure XP beads (Beckman Coulter) on Diagenode IP-Star robot, the size of the DNA was assessed on Fragment Analyzer (Agilent), using the HS Large Fragment 50kb Kit (Agilent). Profiles show the size of lambda DNA before (A) and after treatment with Tagmentase (B).

Figure 2: Fragmentation efficiency depending on the amount of Tagmentase

For fragmentation, 100 ng of DNA from bacteriophage lambda were incubated with Diagenode Tagmentase (Cat. No. C01070010) and Tagmentation buffer (1x) (Cat. No. C01019042) for 7 min at 55°C. The Tagmentase was previously diluted with the Tagmentase Dilution Buffer (Cat. No.) at ¼ and 1/16 dilutions. The reaction was stopped by addition of SDS (0.2% final concentration). After clean-up using AMPure XP beads (Beckman Coulter) on Diagenode IP-Star robot, the size of the DNA was assessed on Fragment Analyzer (Agilent), using the HS Large Fragment 50kb Kit (Agilent). The migration of the samples shows variations of the size distribution according to the amount of Tagmentase used for the reaction.

 Product information

Tagmentase (Tn5 transposase) - unloaded

Protein Molecular weight: 53.3 kDa

Expressed: in Escherichia coli

Product description: Diagenode Tagmentase – unloaded is a hyperactive Tn5 transposase. The enzyme catalyzes “cut and paste” tagmentation reaction and can be used to insert any target DNA in vitro.

Storage conditions: Store at -20°C. Guaranteed stable for 6 months from date of receipt when stored properly.

Storage buffer: Supplied in solution containing 50% v/v glycerol.

Properties & Usage: The enzyme should be loaded with appropriate oligonucleotides prior to use. An efficient transposition require that insert DNA have a specific 19-bp transposase recognition sequence (Mosaic End or ME sequence) at each of its ends. The transposome assembly protocol can be found at https://www.diagenode.com/files/protocols/PRO-Transposome-Assembly-V2.pdf Tagmentase is dependent on Mg++ for activity. Avoid chelators, such as EDTA/EGTA, in reaction buffers. The enzyme is active at pH 7.5-8 at 37-55°C. SDS, EDTA/EGTA or heating to 65°C will inactivate the enzyme.

Applications: Tagmentase (Tn5 transposase) – unloaded can be used in a variety of applications including transgenic experiments, barcoding and library construction for second-generation sequencing. Please note that an additional optimization might be required for custom protocols including the enzyme dose- and time-response experiments.