PaqCI^®

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
R0745S     -20       PaqCI® R0745SVIAL -20 1 x 0.02 ml 10,000 units/ml   rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X   PaqCI Activator S0532SVIAL -20 1 x 0.02 ml 20 µM
R0745L     -20       PaqCI® R0745LVIAL -20 1 x 0.1 ml 10,000 units/ml   rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X   PaqCI Activator S0532LVIAL -20 1 x 0.1 ml 20 µM

Companion Products

• Nuclease-free Water
• NEBridge^® Golden Gate Assembly Kit (BsaI-HF^®v2)

Materials Sold Separately

• rCutSmart™ Buffer

1. PaqCI is an isoschizomer of AarI.
2. This enzyme is impaired by CpG methylation. More information an be found at "Dam-Dcm and CpG Methylation."
3. PaqCI is an enzyme that requires multiple recognition sites to achieve complete digestion.  NEB has therefore included “PaqCI Activator” which can be added to the reaction with 1:1 ratio (i.e. 1 ul enzyme to 1 ul PaqCI Activator).  Please note that for Golden Gate Assembly reactions, the amount of PaqCI Activator varies depending on how many fragments are being assembled. 
4. star activity may result from extended digestion, high enzyme concentration or a glycerol concentration of >5%.

1. Golden Gate Assembly Protocol using PaqCI® (NEB #R0745) and NEBridge Ligase Master Mix (NEB #M1100)
2. Golden Gate Assembly Protocol using PaqCI (NEB #R0745) and T4 DNA Ligase (NEB #M0202)
3. Golden Gate Assembly Protocol for NEBridge® Ligase Master Mix (NEB #M1100)

• Activity of Restriction Enzymes in PCR Buffers
• Cleavage Close to the End of DNA Fragments
• Double Digests
• Effects of CpG Methylation on Restriction Enzyme Cleavage
• Heat Inactivation
• NEBuffer Activity/Performance Chart with Restriction Enzymes
• NEBuffer Comparison Chart (1X)
• Optimizing Restriction Endonuclease Reactions
• Restriction Endonucleases - Survival in a Reaction
• Restriction Enzyme Diluent Buffer Compatibility
• Restriction Enzyme Tips
• Restriction enzymes requiring multi-sites for efficient cleavage
• Single Letter Codes
• Site Preferences
• Star Activity
• Usage Guidelines for Golden Gate Assembly with PaqCI

• Alphabetized List of Recognition Sequences
• Cleavage of Supercoiled DNA
• Dam-Dcm and CpG Methylation
• Enzymes with Nonpalindromic Sequences
• Frequencies of Restriction Sites
• Isoelectric Points (pI) for Restriction Enzymes
• Isoschizomers
• NEB Diluent and Buffer Table
• Time-Saver™ Qualified Enzymes
• Type IIS Restriction Enzymes
• Why Choose Recombinant Enzymes?

• Double Digest Finder
• Enzyme Finder
• NEBridge^® Golden Gate Assembly Tool
• NEBcloner^®
• NEBcutter™ v3.0
• NEBioCalculator®
• REBASE®

1. Which restriction enzymes are used in Golden Gate Assembly?
2. Which restriction enzymes are used in GoldenBraid Assembly?
3. When is star activity a concern?
4. Why is PaqCI not cutting DNA to completion?
5. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?
6. For Golden Gate Assembly, how many base pairs should my amplicon inserts have flanking the PaqCI restriction site?

• Restriction Enzyme 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]

• R0745S_L_v2

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]

• R0745L_v1_10093477
• R0745S_v1_10093476
• R0745L_v1_10117160
• R0745S_v1_10117163
• R0745L_v1_10129863
• R0745S_v1_10129864
• R0745S_v1_10144201
• R0745L_v1_10144203
• R0745S_v1_10155029
• R0745L_v1_10156415
• R0745S_v1_10156417
• R0745L_v1_10165282
• R0745S_v1_10165284
• R0745L_v1_10171799
• R0745S_v1_10171800
• R0745L_v1_10177792
• R0745S_v1_10177790
• R0745S_v1_10177794
• R0745S_v1_10187328
• R0745L_v1_10187329
• R0745L_v1_10198188
• R0745L_v1_10198189
• R0745L_v1_10205489
• R0745S_v1_10198190
• R0745S_v1_10198191
• R0745L_v1_10208036
• R0745L_v2_10211297
• R0745S_v2_10211298
• R0745L_v2_10239410
• R0745S_v2_10238214
• R0745S_v2_10264938
• R0745L_v2_10268659
• R0745S_v2_10285874
• R0745L_v2_10288385
• R0745S_v2_10303967
• R0745S_v2_10309028
• R0745L_v2_10315258
• R0745S_v2_10324196
• R0745L_v2_10309021
• R0745S_v2_10328910
• R0745L_v2_10334635
• R0745L_v2_10342343

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

• PaqCI^®
• rCutSmart™ Buffer

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.