AgeI限制性内切酶使用说明书

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AgeI

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Description

Properties and Usage

Quality Control This enzyme has transitioned to an improved new buffer system. Visit for further details.

The new and current Double Digest Finder and current Activity/Performance Chart for the CutSmart buffer system are available. The previous version of the Double Digest Finder , as well as the previous Version of Activity/Performance Chart that use the former buffer system, are still available for your convenience.

AgeI has High Fidelity (HF) and RE-Mix Master Mix versions available.

Isoschizomers

Catalog #

Size Concentration Price Qty R0552S

300 units 5,000 units/ml $67.001R0552L 1,500 units 5,000 units/ml $268.001

Categories: Restriction Endonucleases: A

Applications: Restriction Enzyme Digestion

Description

Reaction Conditions

1X NEBuffer 1.1

Incubate at 37°C

1X NEBuffer 1.1:

10 mM Bis-Tris-Propane-HCl

10 mM MgCl2

100 μg/ml BSA

pH 7 @ 25°C

Activity in NEBuffers

NEBuffer 1.1: 100%

NEBuffer 2.1: 75%

NEBuffer 3.1: 25%

CutSmart™ Buffer: 75%

Diluent Compatibility

Diluent C

Storage Temperature

-20°C

Storage Conditions

10 mM Tris-HCl

250 mM NaCl

1 mM DTT

0.1 mM EDTA

200 μg/ml BSA

50% Glycerol

0.15% Triton® X-100

pH 7.4 @ 25°C

Heat Inactivation

65°C for 20 min

Methylation Sensitivity

dam methylation: Not Sensitiv e

dcm methylation: Not Sensitiv e

CpG Methylation: Blocked

Activity at Temperature

@37°C: 100%

Quality Control

Quality Control Assays

The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Indiv idual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page. Blue-White Screening (Terminal Integrity):

A sample of DNA v ector linearized with a 10-fold excess of a restriction endonuclease, religated and transformed into an E. coli strain expressing the LacZ beta fragment gene results in less than 1% white colonies.

Exonuclease Activity (Radioactivity Release):

The product is tested in a reaction containing a radiolabeled mixture of single and double-stranded DNA. After incubation for 4 hours the exonuclease activ ity is determined by the % release of radioactiv e nucleotides.

Ligation and Recutting (Terminal Integrity):

After an ov er-digestion of DNA with a restriction endonuclease the percentage of the DNA fragments ligated with T4 DNA ligase and

the percentage that can be recut are determined by agarose gel electrophoresis.

Non-Specific DNase Activity (16 hour):

The product is tested for non-specific nuclease degradation in a reaction containing a DNA substrate. After incubation for 16 hours there is no detectable degradation of the DNA substrate as determined by agarose gel electrophoresis.

Notes

1. Star activ ity may result from extended digestion.

Legal and Disclaimers

Patents

New England Biolabs, Inc.

U.S. Patent No. 7,517,671

Supporting Documents

Certificate of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality control's for an indiv idual lot. The following file naming structure is used to name these document files: [Product Name]COA.