9°N^™ Reverse Gyrase

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
M0200S     -20       9°N™ Reverse Gyrase M0200SVIAL -20 1 x 0.05 ml 5,000 units/ml   Gyrase Reaction Buffer B7636SVIAL -20 1 x 1 ml 5 X

Features

• Introduces positive supercoiling in the presence of ATP
• Enhanced reaction buffer for improved activity
• Thermostable for high temperature workflow applications

Companion Products

• NEBridge^® Golden Gate Assembly Kit (BsmBI-v2)
• NEBridge^® Golden Gate Assembly Kit (BsaI-HF^®v2)
• NEB® PCR Cloning Kit
• T4 DNA Ligase
• Monarch^® Spin Plasmid Miniprep Kit
• NEB® 5-alpha Competent E. coli (High Efficiency)
• NEB® Stable Competent E. coli (High Efficiency)
• T5 Exonuclease

1. The reaction requires ATP and Mg²⁺ (3, 4). The supplied Gyrase Reaction Buffer incorporates these essential components within an optimized formulation, thereby facilitating enhanced gyrase activity.
2. In the absence of ATP, the enzyme will nick circular DNA (5).

1. Lulchev, P., and Klostermeier, D. (2014). NAR. 42 (13), 8200-8213.
2. Thermococcus sp. (strain 9°N-7) isolated by Dr. Holger Jannasch (1991). Woods Hole Oceanographic Institute.
3. Hsieh, TS., and Capp, C. (2005). 208 (21), 20467-20475. JBC.
4. Lee, H. et al. (2025). 53 (12). PubMedID: gkaf581 NAR.
5. Jamroze, A. et al. (2013). 289 (6), 3231-3243. JBC.

1. 9°N™ Reverse Gyrase Protocol (NEB #M0200S)

1. Can DNA Gyrase (E. coli) (NEB #M7636) and 9°N^™ Reverse Gyrase (NEB #M0200) act on linear DNA?
2. Can DNA Gyrase (E. coli) (NEB #M7636) and 9°N^™ Reverse Gyrase (NEB #M0200) create supercoils in both closed and open circular DNA?
3. Are DNA Gyrase (E. coli) (NEB #M7636) and 9°N^™ Reverse Gyrase (NEB #M0200) compatible with transformation or transfection workflows?
4. Are there contaminants that inhibit DNA Gyrase (E. coli) (NEB #M7636) and 9°N^™ Reverse Gyrase (NEB #M0200)?
5. How should DNA treated with DNA Gyrase (E. coli) (NEB #M7636), Topoisomerase I (NEB #M0301), or 9°N^™ Reverse Gyrase (NEB #M0200) be run on a gel?
6. Is the linking number (ΔLk) or the superhelical density (σ) that results from the DNA Gyrase (E.coli) (NEB #M7636) or 9°N^™ Reverse Gyrase (NEB #M0200) reaction fixed or random?
7. Will 9°N^™ Reverse Gyrase (NEB #M0200) add additional positive supercoils in DNA that is already partially supercoiled?
8. Does 9°N^™ Reverse Gyrase (NEB #M0200) work in other NEBuffers?
9. Does 9°N^™ Reverse Gyrase (NEB #M0200) demonstrate additional activities?
10. What is the reaction temperature range for 9°N^™ Reverse Gyrase (NEB #M0200)?
11. Does nicking activity occur at other temperatures for 9°N^™ Reverse Gyrase (NEB #M0200)?
12. What is the difference between DNA Gyrase (E. coli) (NEB #M7636), Topoisomerase I (E. coli) (NEB #M0301), and 9°N^™ Reverse Gyrase (NEB #M0200)?

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]

• M0200S_v1

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

• 9°N^™ Reverse Gyrase

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