This product will be discontinued on September 1, 2016. For superior performance, we recommend you switch to Anza™ 1 NotI.

Not I

Catalog number: 15441041

Invitrogen™  Related applications: Restriction Enzyme Cloning

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Description

5´-GC↓GGCCGC-3´ 3´-CGCCGG↑CG-5´

Invitrogen conventional restriction enzymes come with Universal Buffers L, M, H, K, or T (+BSA). Full details for the this restriction enzymes characteristics are included in the tables below.

Consider upgrading to the Invitrogen™ Anza™ restriction enzyme cloning system for superior convenience. Anza restriction enzymes utilize a single buffer and protocol—complete digestion in 15 minutes at 37°C for all types of DNA substrates. Click here for the Anza equivalent restriction enzyme.
Concentration50 U/ µL
ContentsNot I 10X Buffer H (1mL) 10X Loading Buffer (1mL)
Reaction Temperature37°C
Reaction Mixture Not I 1 µL 10X Buffer H 2 µL 0.10% BSA 2 µL 0.10% Trion X-100 2 µL Substrate DNA <1 µg Sterile water to 20 µL
Heat inactivationEnzyme is inactivated by heating at 65°C for 20 minutes.
Star ActivityUnrelated site may be under conditions of low ionic strength, high enzyme concentration, glycerol concentration >5%, or pH >8.0.
Effect of DNA MethylationEnzyme activity is affected by CG methylase depending on the sequence of the recognition site or the sequence following the site.
Unit DefinitionOne unit is the amount of enzyme required to completely digest 1 µg of λDNA in 50 µL of the reaction mixture in one hour at 37°C.
10X Buffer H500 mM Tris-HCl, pH 7.5 100 mM MgCl2 10 mM Dithiothreitol 1,000 mM NaCl
SourceNocardia otididis-caviarum

Relative Activity in Universal Reaction Buffers

BufferLMHKT(+BSA)
Relative Activity (%)<20*<20*20**<20<20*
*Weak star activity is detected. **100% activity is obtained by addition of 0.01% BSA and 0.01% Triton® X-100.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Enzyme: Not I
Product Size: 1,000 units
Concentration: 50 U⁄µl
Star Activity: May exhibit star activity
Compatible Buffer: 10X Buffer H
Methylation Sensitivity: CpG methylation-sensitive
Optimal Reaction Temperature: 37° C
Sensitive to Heat Inactivation: Yes (Heat-Inactivatable)
Shipping Condition: Approved for shipment on Wet or Dry Ice

Contents & storage

Storage Buffer
10 mM Tris-HCl, pH 7.5
100 mM KCl
0.1 mM EDTA
1 mM Dithiothreitol (DTT)
0.15% Triton® X-100
0.01% BSA
50% (v⁄v) glycerol

Loading Buffer
1% SDS
50% glycerol
0.05% Bromophenol blue

Store enzyme and buffer at -20°C, and loading buffer at RT.

Documents

Manuals & protocols

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