Introducing the Invitrogen™ Anza™ Restriction Enzyme Cloning System, a complete, one-buffer system of restriction enzymes and DNA-modifying enzymes–for beautifully simple cloning.

  • One buffer for all restriction and DNA modifying enzymes
  • One digestion protocol for all DNA types
  • Complete digestion in 15 min
  • Overnight digestion without star activity

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Anza restriction enzyme search tool

Use our Enzyme Search Tool to find the restriction enzymes that best suit your research needs. Search by product name, isoschizomer name, recognition sequence, or SKU number.

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One conductor, a symphony of enzymes

The Anza Restriction Enzyme Cloning System is a complete system of:

128 restriction enzymes
5 DNA-modifying enzymes

All Invitrogen™ Anza™ restriction and DNA modifying enzymes work together cohesively and are fully functional with the single Anza buffer.

The simplicity of Anza restriction enzymes

Anza restriction enzymes are used in a simple two-step protocol, regardless of the number of restriction enzymes in your reaction or the type of DNA you’re using—just prepare your reaction mixture and incubate at 37°C for 15 minutes.

Simple protocol

  1. Prepare a reaction mix by adding reagents in the order indicated in Table 1.
  2. Incubate at 37°C for 15 minutes.

Table 1:

Reagent 1-enzyme 2-enzyme 3-enzyme
Nuclease-Free Water As required to make up final reaction volume
Anza™ Buffer (10X) or Anza™ Red Buffer (10X) 2 µL 2 µL 3 µL
DNA 0.2–1 µg 0.2–1 µg 0.2–1 µg
Anza restriction enzyme 1 1 µL 1 µL 1 µL
Anza restriction enzyme 2 1 µL 1 µL
Anza restriction enzyme 3
1 µL
Final reaction volume 20 µL 20 µL 30 µL

Volumes can be scaled up linearly to 5X.

Simple numbering system

Anza restriction enzymes are named with a combination of the familiar enzyme name and a number.

Restriction enzymes that are more frequently used have a lower number, so you can easily sort, store, and find the enzymes you need, without having to remember more difficult enzyme names when storing alphabetically.    

Sort and store by number

Convenient buffer formats

All Anza restriction enzymes come with an Anza 10X clear buffer and an Anza 10X red buffer to give you the flexibility you require. The red buffer includes a density reagent containing red and yellow tracking dyes that migrate with 800 bp DNA fragments and faster than the 10 bp DNA fragments, respectively, in a 1% agarose gel. This eliminates tedious dye addition steps prior to gel loading and is compatible with downstream applications.*

*For applications that require analysis by fluorescence excitation, Anza 10X buffer is recommended, as the Anza 10X red buffer may interfere with some fluorescence measurements. 

Anza DNA modifying enzymes

Features

Anza restriction enzymes allow for complete digestion in 15 minutes, with one protocol for all DNA types.

Complete digestion of plasmid DNA & PCR product in 15 min

Superior performance


Plasmid DNA expected DNA fragments:
Anza 11 EcoRI, Anza 12 XbaI, Anza 1 NotI – 6,215 bp
Purified PCR product expected DNA fragments:
Anza 11 EcoRI – 1,133 & 506 bp
Anza 12 XbaI – 1,077 & 562 bp
Anza 1 NotI – 1,258 & 381 bp
Map of restriction enzyme cut sites

Figure 1. Anza restriction enzymes are formulated to complete digestion in just 15 minutes. Plasmid DNA (6,215 bp) and purified PCR product (1.6 kb) were digested with Anza restriction enzymes 11 EcoRI, 12 XbaI, and 1 NotI. Reaction mixture followed recommended protocol, which includes 1 µg of DNA and 1 µL of restriction enzyme in a final volume of 20 µL. Incubation was done at 37°C for 15 minutes.

  • L – 1 Kb Plus DNA Ladder
  • C1 – undigested plasmid DNA
  • 1 – Anza 11 EcoRI
  • 2 – Anza 12 XbaI
  • 3 – Anza 1 NotI
  • C2 – undigested PCR fragment
  • 4 – Anza 11 EcoRI
  • 5 – Anza 12 XbaI
  • 6 – Anza 1 NotI

Digestion of plasmid DNA


Expected DNA fragments
Anza 3 BcuI – 5,529 & 686 bp
Anza 47 Eco52I – 4,575, 1,191, & 449 bp
Map of restriction enzyme cut sites

Figure 2. Anza restriction enzymes are formulated to complete digestion in just 15 minutes. Plasmid DNA (6,215 bp) was digested using Anza restriction enzymes 3 BcuI and 47 Eco521, as well as Competitor N SpeI-HF (isoschizomer to BcuI) and Competitor N EagI-HF (isoschizomer to Eco52I). Reaction mixtures included 1 µg of DNA and 1 µL of Anza restriction enzyme to a total volume of 20 µL, following the recommended protocol. Reaction mixtures included 1 µg of DNA and 1 µL of Competitor N restriction enzyme to a total volume of 50 µL, following the manufacturer’s protocol. Incubation was performed at 37°C for 15 minutes for both Anza restriction enzymes and competitor N EagI-HF, as per the protocol. Incubation was done at 37°C for 5 minutes for Competitor N SpeI-HF, per the manufacturer’s protocol.

  • L – 1 Kb Plus DNA Ladder
  • C – undigested plasmid DNA
  • 1 – Anza 3 BcuI
  • 2 – SpeI-HF (Competitor N)
  • 3 – Anza 47 Eco52I
  • 4 – EagI-HF (Competitor N)

Digestion of PCR DNA


Expected DNA fragments
Anza 3 BcuI – 938, 346, 322, & 33 bp
Anza 9 NdeI – 1,486 & 153 bp
Anza 47 Eco52I – 899, 381, 197, & 162 bp
Map of restriction enzyme cut sites

Figure 3. Anza restriction enzymes are formulated to complete digestion in just 15 minutes. Purified PCR product (1.6 kb) was digested using Anza restriction enzymes 3 BcuI, 9 NdeI, and 47 Eco521, as well as Competitor N SpeI-HF (isoschizomer to BcuI), Competitor N NdeI, and Competitor N EagI-HF (isoschizomer to Eco52I). Reaction mixtures included 1 µg of DNA and 1 µL of Anza restriction enzyme to a total volume of 20 µL, following the recommended protocol. Reaction mixtures included 1 µg of DNA and 1 µL of Competitor N restriction enzyme to a total volume of 50 µL, following the manufacturer’s protocol. Incubation was done at 37°C for 15 minutes.

  • L – 1 Kb Plus DNA Ladder
  • C – undigested DNA
  • 1 – Anza 3 BcuI
  • 2 – SpeI-HF (Competitor N)
  • 3 – Anza 9 NdeI
  • 4 – NdeI (Competitor N)
  • 5 – Anza 47 Eco52I
  • 6 – EagI-HF (Competitor N)

Some restriction enzymes can exhibit star activity, or a decrease in specificity to their DNA recognition site, with prolonged digestions. Star activity results in non-specific cleavage of DNA and can occur when reaction conditions are not optimal, such as high glycerol content or presence of Mg2+. Anza restriction enzymes, in conjunction with the Anza buffer, have been optimized to allow for flexibility in digestion times—from complete digestion in 15 minutes to overnight digestions—without the worry of star activity.

Digestion of plasmid DNA and PCR product in 16 hours


Plasmid DNA expected DNA fragments:
Anza 11 EcoRI, Anza 12 XbaI, Anza 1 NotI – 6,215 bp
Purified PCR product expected DNA fragments:
Anza 11 EcoRI – 1,133 & 506 bp
Anza 12 XbaI – 1,077 & 562 bp
Anza 1 NotI – 1,258 & 381 bp
Map of restriction enzyme cut sites

Figure 4. Anza restriction enzymes show no star activity with overnight digestion. Plasmid DNA (6,215 bp) and purified PCR product (1.6 kb) were digested using Anza restriction enzymes 11 EcoRI, 12 XbaI, and 1 NotI. Reaction mixtures included 1 µg of DNA and 1 µL of restriction enzyme to a total volume of 20 µL, following the recommended protocol.  Incubation was done at 37°C for 16 hours.

  • L – 1 Kb Plus DNA Ladder
  • C1 – undigested plasmid DNA
  • 1 – Anza 11 EcoRI
  • 2 – Anza 12 XbaI
  • 3 – Anza 1 NotI
  • C2 – undigested PCR fragment
  • 4 – Anza 11 EcoRI
  • 5 – Anza 12 XbaI
  • 6 – Anza 1 NotI

Anza restriction enzymes allow for complete digestion with multiple enzymes in a single reaction, using the single Anza buffer. Forget the frustrations of trying to find compatible buffers for all your enzymes, and save time with just one protocol for all digestions.

Double digestion


Expected DNA fragments
Anza 47 Eco52I – 4,575, 1,191, & 449 bp
Anza 14 SalI – 4,030 & 2,185 bp
Anza 47 Eco52I + Anza 14 SalI – 2,185, 1,319, 1,191, 1,071, & 449 bp
Map of restriction enzyme cut sites

Figure 5. Anza restriction enzymes show complete digestion with two enzymes in a single buffer. Plasmid DNA (6,215 bp) was digested using Anza restriction enzymes 47 Eco 52I and 14 SalI. Similarly, the same plasmid DNA was digested with Competitor N EagI-HF (isoschizomer to Eco521) and Competitor N SalI-HF. Reaction mixtures included 1 µg of DNA and 1 µL of each Anza restriction enzyme to a total volume of 20 µL, following the recommended protocol. Competitor reaction mixtures included 1 µg of DNA and 1 µL of each Competitor N restriction enzyme to a total volume of 50 µL, per the manufacturer’s protocol.  Incubation was done at 37°C for 15 minutes.

  • L – 1 Kb Plus DNA Ladder
  • C – undigested plasmid DNA
  • 1 – Anza 47 Eco52I
  • 2 – Anza 14 SalI
  • 3 – Anza 47 Eco52I + Anza 14 SalI
  • 4 – EagI-HF (Competitor N)
  • 5 – SalI-HF (Competitor N)
  • 6 – EagI-HF + SalI-HF (Competitor N)

Triple digestion


Expected DNA fragments
Anza 1 NotI – 6,215 bp
Anza 16 HindIII – 6,215 bp
Anza 15 XmaJI – 6,215 bp
Anza 1 NotI + Anza 16 HindIII + Anza 15 XmaJI – 4,285, 1,075, & 855 bp
Map of restriction enzyme cut sites

Figure 6. Anza restriction enzymes show complete digestion with three enzymes in a single buffer. Plasmid DNA (6,215 bp) was digested using Anza restriction enzymes 1 NotI, 16 HindIII, and 15 XmaJI. For single restriction enzyme digestions, reaction mixture included 1 µg of DNA and 1 µl of restriction enzyme to a total volume of 20 µL.  Reaction mixtures included 1 µg of DNA and 1 µL of each restriction enzyme to a total volume of 30 µL for triple digestion, per the recommended protocol.  Incubation was done at 37°C for 15 minutes.

  • C – undigested plasmid DNA
  • 1 – Anza 1 NotI
  • 2 – Anza 16 HindIII
  • 3 – Anza 15 XmaJI
  • 4 – Anza 1 NotI + Anza 16 HindIII + Anza 15 XmaJI
  • L – 1 Kb Plus DNA Ladder

Anza starter kits



Order now ›
 
Starter kit contains conc (U/µl) volume (µl) Units (U)
Anza 1 NotI 20 20 400
*Anza 3 BcuI
10 20 200
Anza 5 BamHI 10 20 200
Anza 8 XhoI
20 20 400
Anza 10 DpnI 20 20 400
Anza 11 EcoRI 20 20 400
Anza 12 XbaI
10 20 200
Anza 16 HindIII 20 20 400
Anza 19 BglII 10 20 200
**Anza 26 Eco32I 20 20 400
Anza T4 DNA Ligase Master Mix 4x 100 20 rxns
Anza Alkaline Phosphatase 1 20 20 rxns
Anza 10X Buffer 10x 500 n/a
Anza 10X Red Buffer 10x 500 n/a

*Isoschizomer to prototype SpeI
**Isoschizomer to prototype EcoRV

An isoschizomer is a protein that recognizes the same recognition sequence and digests the DNA in the same location & pattern.


Order now ›
 
Starter kit contains conc (U/µl) volume (µl) Units (U)
Anza 1 NotI 20 20 400
Anza 5 BamHI 10 20 200
Anza 11 EcoRI
20 20 400
Anza 12 XbaI 10 20 200
Anza 16 HindIII
20 20 400
Anza T4 DNA Ligase Master Mix 4x 100 20 rxns
Anza Alkaline Phosphatase 1 20 20 rxns
Anza 10X Buffer 10x 500 n/a
Anza 10X Red Buffer 10x 500 n/a

Download the CloningBench 3.0 mobile App

CloningBench is designed to provide you with useful and essential tools to help guide your cloning experiments at the bench or on the move. Find the right Anza enzyme for your research and order directly using the CloningBench mobile app. Download for free for iOS™ and Android™ devices today.

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Resources

NEW Invitrogen ˜­Anza Restriction Enzymes for Uploading to Vector NTI

Instructions for Importing Anza Restriction Enzymes
Vector NTI Express or Vector NTI Express Designer
Vector NTI Advance

Data File for Importing
 Anza Restriction Enzyme Data File