28S- 300F/1846alt 20100129

Basic information

Date: 
Thu, 2010-01-28 23:00
Gene: 
Template: 
dna

Procedure

PCR cocktail: 
Component Volume (ul) Mix (ul)
Forward primer  0.5  
Reversed primer  0.5  
Buffer    
MgCl2    
dNTPs    
Taq    
DNA (if nested)  2  
ddH2O
 22  
Total volume  25
Forward primer(s): 
Thermal cycling (abbreviated): 
95ºC@4min, 2X(94ºC@30sec,56-54ºC@30sec, 72ºC@60sec)36x(94ºC@30sec, 52ºC@30sec, 72ºC@60sec), 72ºC@10min
PCR machine: 
GeneAmp
Thermal cycling: 
Step Comment Cycles Time (sec) Temperature (°C)
Initialization For DNA polymerases that need hot-start heat activation 1
240
95  
Denaturation DNA melting producing single-stranded DNA      
Amplification   2 -  -
Denaturation DNA melting producing single-stranded DNA
 - 30 94 
Annealing The primer binds to the template  - 30 56-54
Extension The primer binds to the template
 - 60  72
Amplification   36    
Denaturation     30 94
Annealing     30 52
Extension     60 72
Final elongation Any remaining single-stranded DNA is extended 1 600  72
Hold   1   12
File attachments: 
Products and Equipment: 

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Scratchpads developed and conceived by (alphabetical): Ed Baker, Katherine Bouton Alice Heaton Dimitris Koureas, Laurence Livermore, Dave Roberts, Simon Rycroft, Ben Scott, Vince Smith