FastDigest NcoI

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FastDigest NcoI

5' C ↓C A T G G 3' 3' G G T A C ↑C 5' Thermo Scientific FastDigest NcoIRead more

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Catalog Number	Quantity
FD0574	100 μL (100 Reactions)
FD0575	300 μL (300 Reactions)
FD0573	20 μL (20 Reactions)

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Catalog number FD0574

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100 μL (100 Reactions)

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

Thermo Scientific FastDigest NcoI restriction enzyme recognizes C^CATGG site and cuts best at 37°C in 5–15 minutes using universal FastDigest Buffer. Isoschizomers: Bsp19I.

Thermo Scientific FastDigest NcoI 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 available

Applications

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

Note: 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.

Specifications

Compatible Buffer10X FastDigest Buffer/FastDigest Green Buffer

Inactivation Temperature65°C

Overhang5' protruding ends

Product TypeRestriction Enzyme

Quantity100 μL (100 Reactions)

EnzymeNco I

Methylation SensitivityNot Dam Methylation-Sensitive, Not Dcm Methylation-Sensitive, Not CpG Methylation-Sensitive

Optimal Reaction Temperature37°C

Product LineFastDigest™

Research CategoryTraditional Cloning

Sensitive to Heat InactivationYes

Type IIS RENo

Unit SizeEach

Contents & Storage

• FastDigest NcoI (100 μL)
• 10X FastDigest Buffer (1.0 mL)
• 10X FastDigest Green Buffer (1.0 mL)

Store at –20°C.

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Frequently asked questions (FAQs)

Can I store the FastDigest Buffers at 4 degrees C?

The FastDigest Buffers can be stored at 4 degrees C for at least 6 months. For long-term storage until their expiration, we recommend storing the FastDigest and FastDigest Green Buffers at -20 degrees C.

How can I perform large scale digestion with the FastDigest restriction enzymes?

Our general recommendation is to use 5-10 µL of FastDigest enzymes with 100 µg DNA in 500 µL and 10-20 µL of the FastDigest enzymes with 200 µg DNA in 1 mL. Incubate the reaction for 16 h (or the longest recommended incubation time if the particular enzyme is prone to star activity) at the recommended temperature. Optimal conditions may vary as complete digestion of DNA depends on the nature of the particular FastDigest restriction enzymes and the DNA substrates.

How I can transform or calculate reaction numbers in the unit of FastDigest enzymes?

The concentration of FastDigest enzymes is proprietary information. The FastDigest enzyme activity is measured by FastDigest unit (FDU): 1 µL of enzyme (1 FDU) cleaves 1 µg of DNA substrate in 5-15 min at 37 degrees C in 20 µL of 1X FastDigest buffer.

Are the FastDigest and FastDigest Green Buffer available separately?

Yes, the FastDigest Buffer (Cat. No. B64) and the FastDigest Green Buffer (Cat. No. B71) can be purchased separately.

What is the difference between FastDigest Buffer and FastDigest Green Buffer?

The FastDigest Green Buffer offers the same performance as the colorless FastDigest Buffer but enables direct loading of the reaction mixture on gels. The 10X FastDigest Green Buffer includes a density reagent and two tracking dyes for direct loading. The blue dye migrates with 3-5 kb DNA fragments in a 1% agarose gel and has an excitation peak of 424 nm. The yellow dye migrates faster than 10 bp DNA fragments in a 1% agarose gel and has an excitation peak of 615 nm.

Find additional tips, troubleshooting help, and resources within our Restriction Enzyme Cloning Support Center.

View all FastDigest NcoI, 100 μL (100 Reactions) FAQs