Product Class: Restriction Endonuclease

ApaI
neb31 cloned at NEB recombinant time saver dil_A 37 65 Heat CpG dcm

New 37°C incubation temperature.

ApaI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10137117. All subsequent (higher number) lots will contain rAlbumin. For additional information, please visit at Learn more.

We are excited to announce that all reaction buffers are now BSA-free. NEB began switching our BSA-containing reaction buffers in April 2021 to buffers containing Recombinant Albumin (rAlbumin) for restriction enzymes and some DNA modifying enzymes. Find more details at www.neb.com/BSA-free.

NEB restriction endonuclease that recognizes the sequence G_GGCC^C
Isoschizomers | Single Letter Code

Product Introduction

Catalog # Size Concentration
R0114S 5000.0 units 50000 units/ml
R0114L 25000.0 units 50000 units/ml

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Product Information

Description

Product Source

An E. coli strain that carries the ApaI gene from Acetobacter pasteurianus sub. pasteurianus(ATCC 9432).
This product is related to the following categories:
Restriction Endonucleases: A Products,
Time-Saver Qualified Restriction Enzymes Products,
This product can be used in the following applications:
Fast Cloning: Accelerate your cloning workflows with reagents from NEB,
Restriction Enzyme Digestion

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to digest 1 µg of pXba DNA in 1 hour at 37°C in a total reaction volume of 50 µl.

Reaction Conditions

1X rCutSmart™ Buffer
Incubate at 37°C

1X rCutSmart™ Buffer
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
100 µg/ml Recombinant Albumin
(pH 7.9 @ 25°C)

Activity in NEBuffers

NEBuffer™ r1.1: 25%
NEBuffer™ r2.1: 25%
NEBuffer™ r3.1: <10%
rCutSmart™ Buffer: 100%

Diluent Compatibility

Storage Buffer

10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
500 µg/ml Recombinant Albumin
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

65°C for 20 minutes

Methylation Sensitivity

dam methylation: Not Sensitive
dcm methylation: Blocked by Overlapping
CpG Methylation: Blocked by Overlapping

Isoschizomers

Bsp120I
PspOMI

Activity at Temperature

@25°C: 100%

Product Notes

  1. ApaI is inhibited by salt concentrations above 50 mM.
  2. Blocked by overlapping dcm methylation and overlapping CpG methylation.

Protocols, Manuals & Usage

Protocols

  1. Optimizing Restriction Endonuclease Reactions
  2. Protocol for Direct Digestion of gDNA during droplet digital PCR (ddPCR)
  3. Restriction Digest Protocol
  4. Protocol for Direct Digestion of gDNA during droplet digital PCR (ddPCR)
  5. Double Digest Protocol with Standard Restriction Enzymes

Usage & Guidelines

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. Is ApaI activity sensitive to dam, dcm or mammalian CpG methylation?
  2. How many base pairs should be added at the end of a PCR primer after the ApaI recognition site to guarantee that ApaI will cut properly?
  3. Does ApaI have any neoschizomers?
  4. It seems that ApaI is having some difficulties cutting my DNA. Is there a reason for that?
  5. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
  6. I tested your restriction enzyme on the substrate DNA recommended by NEB, and it appears to be active, however it does not digest my DNA. What could be the reason?
  7. Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?
  8. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?
  9. In an effort to standardize reaction conditions, the recommended incubation temperature has changed for ApaI (NEB #R0114), BaeI (NEB #R0613), BsrDI (NEB #R0574) and SmaI (NEB #R0141). Which incubation temperature should I use?
  10. Are certain restriction enzymes more active at higher incubation temperatures than what is recommended?

Troubleshooting

Quality, Safety & Legal

Quality Assurance Statement

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specification Change Notifications

ApaI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10137117. All subsequent (higher number) lots will contain rAlbumin. For additional information, please visit at NEB Restriction Enzyme formulations with Recombinant Albumin (rAlbumin).

Effective 3/7/2022, new 37°C incubation temperature.

 

Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Certificate Of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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