The Stylet

Diversity and Systematics of Acoela and Nemertodermatida

18S- S30/5FK 20081002

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, 40x(94ºC@30sec, 52ºC@30sec, 72ºC@30sec), 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   40    
Denaturation     30 94
Annealing     30 52
Extension     30 72
Final elongation Any remaining single-stranded DNA is extended 1 600  72
Final hold Short-term storage 1 - 12 
Notes: 
2 worms were extracted separately from sample tube UJ08-119. UJ08-119 (no. 11 ) first extraction. UJ08-119-alt (no.12) second extraction.
PCR program: 
S30
Reversed primer(s): 
File attachments: 
Products and Equipment: 

Add new comment

To prevent automated spam submissions leave this field empty.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Image CAPTCHA
Enter the characters shown in the image.
Scratchpads developed and conceived by (alphabetical): Ed Baker, Katherine Bouton Alice Heaton Dimitris Koureas, Laurence Livermore, Dave Roberts, Simon Rycroft, Ben Scott, Vince Smith