The Stylet

Diversity and Systematics of Acoela and Nemertodermatida

28S- 1200F/2450R 20071107

Basic information

Date: 
Tue, 2007-11-06 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) 1  
ddH2O
 22  
Total volume  24
Forward primer(s): 
Thermal cycling (abbreviated): 
95ºC@5min, 40x(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  
 
Denaturation DNA melting producing single-stranded DNA 1 300 95 
 Amplification   40    
 Denaturation     30 94
 Annealing     30 52
 Extension     60 72
Final elongation Any remaining single-stranded DNA is extended 1 600  72
Hold   1   8
Reversed primer(s): 
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