Team:HokkaidoU Japan/Notebook/Length/as90 120

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Anti-sense 90, 120 Construction

    
      
        
	  
	  Start
	  
	

	  
	  
PCR & PCR purification

	  
	  
	    
	      
	        
		  
Fri Oct 3, Mon Oct 6

		
	      
	      
	R0040-B0034-E1010-B0015 as RBS XhoI, 90bp as mRFP Nco as RBS XhoI, 120bp as mRFP Nco

	    
	  
	

	  
	  
Digestion

	  
	  
	    
	      
	        
		  
Sat Oct 4

		
	      
	      
Cut as 90, as120, H-stem with NcoI, XhoI(using 10×Cut Smart)

	    
	  
	

	  
	  
Ethanol precipitation

	  
	  
	    
	      
	        
		  
Sat Oct 4

		
	      
	      
as90, as120

	    
	  
	

	  
	  
Gel Extraction

	  
	  
	    
	      
	        
		  
Sat Oct 4

		
	      
	      
H-stem

	    
	  
	

	  
	  
Dephophorylation

	  
	  
	    
	      
	        
		  
Sat Oct 4

		
	      
	      
H-stem

	    
	  
	

	  
	  
Ligation

	  
	  
	    
	      
	        
		  
Sun Oct 5

		
	      
	      
Ligate as90, as120 with H-stem

	    
	  
	

	  
	  
Transformation

	  
	  
	    
	      
	        
		  
Sun Oct 5

		
	      
	      
as90, as120 on H-stem 5µL DNA to DH5α Turbo

	    
	  
	

	  
	  
Colony PCR

	  
	  
	    
	      
	        
		  
Mon Oct 6

		
	      
	      
as90, as120 on H-stem

	    
	  
	

	  
	  
Liquid Culture

	  
	  
	    
	      
	        
		  
Tue Oct 7

		
	      
	      
as90, as120 on H-stem

	    
	  
	

	  
	  
Mini-prep

	  
	  
	    
	      
	        
		  
Wed Oct 8

		
	      
	      
as90, as120 on H-stem

	    
	  
	

	  
	  
Dye terminator

	  
	  
	    
	      
	        
		  
Wed Oct 8

		
	      
	      
as90, as120 on H-stem

	    
	  
	
	
      Complete!