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Anza Restriction Enzymes
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
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.
Browse Anza enzymes View the Anza Type IIS restriction enzyme white paper
One conductor, a symphony of 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
- Prepare a reaction mix by adding reagents in the order indicated in table below.
- Incubate at 37°C for 15 minutes.
| 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
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
- Prepare a reaction mix by adding reagents in the order indicated in table below.
- Incubate at 37°C for 15 minutes.
| 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.
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 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.
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
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*Isoschizomer to prototype SpeI |
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