The Stylet

Diversity and Systematics of Acoela and Nemertodermatida

28S- UJ2176/3449L 20080617

Basic information

Date: 
Mon, 2008-06-16 22:00
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,58-54ºC@30sec, 72ºC@60sec),34X (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
 
Denaturation DNA melting producing single-stranded DNA 1 240 95 
 Amplification   2    
 Denaturation     30 94
      30 58-54
      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
Reversed primer(s): 
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