DNA Gyrase Assay Kit | Gyrase Drug Screening Kit | 고마바이오텍

• Gyrase 절단을 유도하는 interfacial poison (IFP) 및 Gyrase 작용을 차단하는 Catalytic inhibitory compound (CIC)와 같은 Gyrase 표적 약물을 스크리닝 하도록 설계되어 있습니다. 
• DNA Gyrase의 DNA supercoliling action이 relaxed plasmid DNA substrate (pHOT1)에 어떻게 작용하는지 체크함으로써  drug의 효과를 평가할 수 있습니다.

구성 

• Relaxed pHOT1 DNA (50 ug total)  
• Supercoiled pHOT1 DNA (25 ul in gel loading buffer).  Load 2 ul as a marker.
• Linearized pHOT1 DNA (25 ul in gel loading buffer).  Load 2 ul as a marker for cleavage (DS DNA breaks).
• 5x Gyrase assay buffer (600 ul) 
• Dilution buffer (600 ul).  Dilute gyrase with this buffer.  
• 10x gel loading buffer (300 ul): 0.25% bromophenol blue, 50% glycerol, use 0.1 volume in reactions.
• 10x Proteinase K (100 ul) at 0.5 mg/ml (10x stock of proteinase K and should be diluted to a final 1x to degrade proteins after reaction termination; i.e. use at 50 ug/mL final).
• 10% SDS (300 ul).  To be added to reaction mix to give a final concentration of 1% to terminate the relaxation reactions.
• Purified DNA gyrase; 100 units 
• Ciprofoxacin 10ug/ml:  50 ul (10x Stock; dilute 1:10 in final reaction mix to give 1 ug/ml final).