Team:Arizona State/Lab/Protocols/PCR

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{{:Team:Arizona State/Templates/main|title=[[Team:Arizona State/Lab/Protocols|Protocols]]: PCR|content=
{{:Team:Arizona State/Templates/main|title=[[Team:Arizona State/Lab/Protocols|Protocols]]: PCR|content=
 
 
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'''PCR:'''
 
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----
 
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* Prepare master mix (for 8 tubes):
 
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* 10*n uL 5x Phusion buffer
 
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* 4*n uL 2.5 mM dNTPs
 
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* 2.5*n uL forward primer
 
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* 2.5*n uL reverse primer
 
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* 29.5*n uL PCR water
 
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* Add 48 uL master mix to each of seven tubes.
 
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* Label tubes blank, 0, 1, 2, 4, 8, 10
 
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* Add DNA and MgCl2 to tubes according to chart:
 
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<!-- it appears that tables don't work due to the piping of content-->
 
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Blank 0 1 2 4 8 10<br>
 
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MgCl2 0 0 .5 1 2 4 5<br>
 
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DNA  0 1 1 1 1 1 1<br>
 
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* Add 0.5 uL Phusion polymerase to each tube.
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# For 50 ul reactions, prepare master mix as follows for '''n''' tubes:
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* Place tubes in thermocycler.
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#: 10*n uL 5x Phusion buffer
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* Adjust settings as needed.
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#: 4*n uL 2.5 mM dNTPs
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#: 2.5*n uL forward primer
 +
#: 2.5*n uL reverse primer
 +
#: 29.5*n uL PCR water
 +
# Add 48 uL master mix to each of seven tubes.
 +
# Label tubes blank, 0, 1, 2, 4, 8, 10
 +
# Add DNA and MgCl2 to tubes according to chart:
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<center>{{:Team:Arizona State/Templates/PCR table}}</center>
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# Add 0.5 uL Phusion polymerase to each tube.
 +
# Place tubes in thermocycler.
 +
# Adjust settings as needed.
}}
}}

Latest revision as of 23:01, 26 September 2011


Protocols: PCR


ASU Logo.png
 
  1. For 50 ul reactions, prepare master mix as follows for n tubes:
    10*n uL 5x Phusion buffer
    4*n uL 2.5 mM dNTPs
    2.5*n uL forward primer
    2.5*n uL reverse primer
    29.5*n uL PCR water
  2. Add 48 uL master mix to each of seven tubes.
  3. Label tubes blank, 0, 1, 2, 4, 8, 10
  4. Add DNA and MgCl2 to tubes according to chart:
blank 0 1 2 4 8 10
MgCl2 0 0 .5 1 2 4 5
DNA 0 1 1 1 1 1 1
  1. Add 0.5 uL Phusion polymerase to each tube.
  2. Place tubes in thermocycler.
  3. Adjust settings as needed.