5'

↓

N7

G

A

A

C

N6

T

C

N td>

C

N12-13

↓

3'

3'

↑

N12-13

C

T

T

G

N6

A

G

G

N7

↑

5'

Thermo Scientific AloI restriction enzyme recognizes^(7/12 -13)GAAC(N)6TCC(12-13/7)^ site, and the best cleavage effect is in R buffer at 30°C. For a table of enzymatic activities, double digestion conditions, and heat inactivation of this and other restriction enzymes, see Reaction conditions for restriction enzymes.

Thermo Scientific Conventional Restriction Endonucleases are a large collection of high quality restriction enzymes optimized to function in one buffer of a five-buffer system. In addition, a universal Tango buffer is provided to facilitate double enzyme digestion. All enzymes demonstrated 100% activity under recommended buffers and reaction conditions. To ensure consistent performance, Thermo Scientific Restriction Enzyme Reaction Buffers contain premixed BSA, which enhances the stability of multiple enzymes and binds contaminants that may be present in DNA preparations.

Features

• Superior quality—strict quality control and industry-leading production processes
• Convenient color-coded five-buffer system
•Includes universal Tango buffer for double digestion
•Pre-mixed BSA in reaction buffer
•Multiple restriction endonuclease specificities available

< b>Applications

•Molecular cloning
•Restriction site mapping
•Genotyping
•Southern blot
•Restriction fragment length polymorphism ( RFLP)
•SNP

Notes: See product specifications for methylation sensitivity. 20% active when incubated at 37°C. AloI cuts double-stranded DNA on both sides of the recognition site, forming a double-stranded nick. Its unique feature is that it can generate a degenerate restriction site at the 3-end of the recognition sequence (at the 12th or 13th nucleotide). Adding SAM to the reaction mixture will result in incomplete AloI digestion. Over-digestion with more than 10-fold AloI may result in asterisk activity. AloI may bind to severed DNA. This can cause DNA band drift during electrophoresis. To avoid atypical DNA banding patterns, use 6X DNA loading dye with SDS solution for sample preparation, or heat digested DNA in the presence of SDS before electrophoresis.