SphI Restriction Enzyme

**Rapid-digestion restriction enzymes** are engineered restriction enzymes designed for fast DNA digestion. These enzymes show strong activity in a **universal digestion buffer** and can digest DNA within **5–15 minutes**. This enables different combinations of restriction enzymes to work simultaneously in a single reaction tube and reduces the need for sequential digestion steps.

Rapid-digestion restriction enzymes undergo strict quality control testing and can be used to digest **plasmid DNA, genomic DNA, viral DNA, and PCR products**. The **color loading digestion buffer** contains a density reagent together with red and yellow tracking dyes, allowing reaction mixtures to be loaded directly onto an agarose gel after digestion. In a **1% agarose gel**, the red dye migrates similarly to approximately **2.5 kb double-stranded DNA fragments**, while the yellow dye migrates similarly to approximately **10 bp double-stranded DNA fragments**.

Restriction Endonucleases

Rapid-Digestion Restriction Endonucleases

The reaction can be completed within 15 minutes Recommended Reaction Conditions 1x CutOne® Buffer; Incubate at 37°C; Refer to "Protocol for Fast DNA Digestion" for reaction setup. Heat Inactivation Incubation at 80°C for 20 minutes. Methylation sensitivity Cleavage with this restriction enzyme may be blocked or impaired when the substrate DNA is methylated by the EcoBI methylase.

20 U/μL

GCATGC 5'...G C A T G ↓ C...3' 3'...C ↑ G T A C G...5'

PaeI

• Rapid-digestion SphI (20 U/μl): 50 μl / 1,000 U
• 10× Universal Digestion Buffer: 1 ml
• 10× Color Loading Digestion Buffer: 1 ml

-20°C

2~8°C