Takara QuickCut restriction enzymes

Perform restriction digestion of DNA in just 5 minutes using any of our QuickCut enzymes. Each QuickCut restriction enzyme is formulated to allow complete digestion of linear DNA, plasmid DNA, or PCR products in 5 to 30 minutes. Each QuickCut restriction enzyme product is supplied with both 10X QuickCut Buffer and 10X QuickCut Green Buffer. Save time with easy-to use, fast-digesting QuickCut enzymes.

Overview

• Allows fast digestion of DNA (5 min for most QuickCut enzymes and DNA types)
• Supplied with 10X QuickCut Buffer and 10X QuickCut Green Buffer
• 10X QuickCut Green Buffer allows direct loading of the reaction on an agarose gel

More Information

Applications

• Fast preparation of DNA inserts for cloning
• Rapid clone analysis
• RFLP genotyping
• Analysis of PCR products by fast restriction enzyme digestion

10X Buffers

10X QuickCut Buffer does not contain loading dye and may be used for applications that are hindered by fluorescence excitation. Use this colorless buffer when analyzing reactions by detection of fluorescence signal.

10X QuickCut Green Buffer contains loading buffer for direct application of restriction reactions to agarose gels, which simplifies sample handling and minimizes pipetting. Two tracking dyes are included: a blue dye that migrates at the location of 3–5-kb DNA fragments in a 1% agarose gel, and a yellow dye that migrates at the location of 100-bp DNA fragments in a 1% agarose gel.

Components

• QuickCut restriction enzyme (50 rxns, 50 μl)
• 10X QuickCut Buffer (500 μl)
• 10X QuickCut Green Buffer (500 μl)

Storage

–20°C

Specifications

For restriction enzyme recognition sites, effects of DNA methylation, enzyme source organism, and more information, refer to the Data Sheet for each QuickCut restriction enzyme. Click on the Resources tab to view technical literature.

Standard QuickCut digestion protocol

1. Prepare reactions according to the following table

   	Linear DNA	Plasmid DNA	PCR product
   10X QuickCut Buffer or 10X QuickCut Green Buffer	5 µl	5 µl	3 µl
   DNA	Up to 1 µg	Up to 1 µg	Up to 0.2 µg
   QuickCut enzyme	1 µl	1 µl	1 µl
   Sterile water	To a final volume of 50 µl	To a final volume of 50 µl	To a final volume of 30 µl

2. Mix gently and centrifuge briefly.

3. Incubate at the recommended reaction conditions for each QuickCut enzyme. (Check each product’s Data Sheet for recommended incubation temperature and length. Incubate BamH I, Cla I, and Sma I reactions at 30°C. For others, incubate at 37°C.)

Quality control assays

• Functional Test
  To verify activity, 1 µl QuickCut restriction enzyme is verified to completely digest 1 µg of linear DNA with 1 µl QuickCut restriction enzyme in a 50-µl reaction at the recommended temperature and time for each enzyme.
• Star Activity Test
  To check for star activity, 1 µl of QuickCut restriction enzyme is used to digest 1 µg DNA for 16 hours at the recommended temperature for each enzyme. The reaction is subjected to DNA electrophoresis to confirm that no changes occur in the band pattern.
• Labeled Oligonucleotide Assay (LOA) Test
  1 µl of QuickCut restriction enzyme is incubated with fluorescently labeled oligonucleotide for 1 h at the recommended incubation temperature for each enzyme. The sample is analyzed to ensure that the decomposition rate of the labeled oligonucleotide is less than 10%.

Precautions

• To avoid star activity, do not digest for more than 16 hr.
• The total volume of restriction enzyme should not exceed 1/10 of the reaction volume.
• When performing double digestion with two enzymes that require different reaction temperatures, perform a first digestion with the QuickCut enzyme requiring a lower reaction temperature, then add the second QuickCut enzyme and shift to the higher reaction temperature required by the second enzyme.

Additional product information

Please see the product`s Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.