|5'|| || ||↓||N7||G ||A ||A ||N7||T ||T ||G ||G ||N11||↓|| ||3'|
|3'|| || ||↑||N12||C ||T ||T ||N7||A ||A ||C ||C ||N6||↑|| ||5'|
Thermo Scientific FastDigest AjuI restriction enzyme recognizes ^(7/12)GAA(N)7TTGG(11/6)^ site and cuts best at 37°C in 5–15 minutes using universal FastDigest Buffer.
Thermo Scientific FastDigest AjuI is one of an advanced line of fast restriction enzymes that are all 100% active in the universal FastDigest and FastDigest Green reaction buffers.
The universal buffer allows rapid single-, double-, or multiple DNA digestion within 5–15 minutes eliminating any need for buffer change or subsequent DNA clean-up steps. See Reaction Conditions for FastDigest Enzymes
for a table of enzyme activity, heat inactivation and incubation times for this and other FastDigest restriction enzymes. DNA modifying enzymes, such as Klenow Fragment
, T4 DNA Ligase
, alkaline phosphatases and T4 DNA Polymerase
all have 100% activity in FastDigest Buffer. Therefore, enzymes for downstream applications can be directly added to the FastDigest reaction mix.
For additional convenience FastDigest Green Buffer includes a density reagent and two tracking dyes for direct loading of digestion reaction products on gels.
Short incubation times and optimal composition of the universal FastDigest Buffer eliminate star activity effects.Features
• 100% activity of all FastDigest enzymes in the universal buffer
• 100% buffer compatibility with downstream applications
• Complete digestion in 5–15 minutes
• Direct loading on gels
• No star activity
• 176 FastDigest enzymes availableApplications
• Molecular cloning
• Restriction site mapping
• Southern Blot
• Restriction fragment length polymorphism (RFLP)
• SNP analysisNote:
FastDigest AjuI requires SAM for activity. Complete cleavage of some substrates with FastDigest AjuI is difficult to achieve. FastDigest AjuI produces double-strand cuts on both sides of the interrupted recognition site. The exact cleavage position depends on the sequences flanking the recognition site and may shift by one base pair. For an individual sequence, there will be a mix of the 2 cleavage positions with the majority of the digestion occurring at one of the positions.
The FastDigest Green Buffer offers the same high performance in DNA digestion and downstream applications as the colorless FastDigest Buffer. For applications that require product analysis by fluorescence excitation (e.g. concentration measurements in UV light) the colorless FastDigest Buffer is recommended.
For methylation sensitivity, refer to product specifications.
For Research Use Only. Not for use in diagnostic procedures.