EnGen^® Mutation Detection Kit

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
E3321S     -20       Q5® Hot Start High-Fidelity 2X Master Mix M0494AAVIAL -20 1 x 0.4 ml 2 X   NEBuffer™ 2 B7002SVIAL -20 1 x 1.25 ml 10 X   Control Template and Primer Mix S0534AVIAL -20 1 x 0.025 ml 10 X   Proteinase K, Molecular Biology Grade P8107AVIAL -20 1 x 0.12 ml 800 units/ml   Quick-Load® Purple 1 kb Plus DNA Ladder N0550AVIAL 4 1 x 0.2 ml 100 µg/ml   Gel Loading Dye, Purple (6X), no SDS B7025SVIAL 25 1 x 1 ml 6 X   EnGen® T7 Endonuclease I M0450AVIAL -20 1 x 0.025 ml Not Applicable

Companion Products

• Cas9 Nuclease, S. pyogenes
• Gel Loading Dye, Purple (6X), no SDS
• Quick-Load® Purple 1 Kb Plus DNA Ladder
• Q5® Hot Start High-Fidelity 2X Master Mix
• Monarch® PCR & DNA Cleanup Kit (5 μg)
• HiScribe^®; T7 Quick High Yield RNA Synthesis Kit
• NEBuilder® HiFi DNA Assembly Cloning Kit
• NEBuilder® HiFi DNA Assembly Master Mix
• EnGen® sgRNA Synthesis Kit, S. pyogenes

1. T7 Endonuclease I-based Mutation Detection with the EnGen^® Mutation Detection Kit (NEB #E3321)
2. Transfection of Cas9 RNP (ribonucleoprotein) into adherent cells using the Lipofectamine® RNAiMAX

• Determining Efficiency of On-Target CRISPR Cas9 Genome Editing Using the NEB^® EnGen^® Mutation Detection Kit on LabChip Gel Xpress

1. Why do I see an extra band when I run the undigested heteroduplex on an agarose gel?
2. Why do I see very little DNA on my gel or low signal on the fragment analyzer?
3. Will EnGen^® T7 Endonuclease I recognize single base mismatches and single base insertions or deletions (indels)?  
4. Can cell lysates be used in the PCR reaction?
5. What are the expected results using the included control?
6. My PCR reaction yield is low, can I add more than 5 µl of the PCR reaction to the digestion reaction?

• PCR Troubleshooting Guide

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.

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]

• E3321S_v1

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]

• E3321S_v1_10039773
• E3321S_v1_10058712
• E3321S_v1_10064780
• E3321S_v1_10069545
• E3321S_v1_10081167
• E3321S_v1_10092905
• E3321S_v1_10097239
• E3321S_v1_10106417
• E3321S_v1_10111218
• E3321S_v1_10115975
• E3321S_v1_10122065
• E3321S_v1_10130231
• E3321S_v1_10136978
• E3321S_v1_10144368
• E3321S_v1_10156034
• E3321S_v1_10162352
• E3321S_v1_10166683
• E3321S_v1_10170600
• E3321S_v1_10179186
• E3321S_v1_10191099
• E3321S_v1_10203706
• E3321S_v1_10212533
• E3321S_v1_0041711
• E3321S_v1_10217191
• E3321S_v1_10233004
• E3321S_v1_10241665

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

• Q5^® Hot Start High-Fidelity 2X Master Mix
• NEBuffer™ 2
• Control Template and Primer Mix
• Proteinase K, Molecular Biology Grade
• Quick-Load^® Purple 1 kb Plus DNA Ladder
• Gel Loading Dye, Purple (6X), no SDS
• EnGen^® T7 Endonuclease I

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.

Licenses

This product is covered by one or more patents.

This product is licensed for research and commercial use from Bio-Rad Laboratories, Inc., under U.S. Pat. Nos. 6,627,424, 7,541,170, 7,670,808, 7,666,645, and corresponding patents in other countries. No rights are granted for use of the product for Digital PCR or real-time PCR applications, with the exception of quantification in Next Generation Sequencing workflows.

For additional information or to inquire about commercial use, please contact busdev@neb.com.