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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{| border="1" cellspacing="0" cellpadding="2" bordercolor="black"
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# For 50 ul reactions, prepare master mix as follows for '''n''' tubes:
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|+ optional table caption
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#: 10*n uL 5x Phusion buffer
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! Column heading 1 !! Column heading 2 !! Column heading 3
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#: 4*n uL 2.5 mM dNTPs
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|-
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#: 2.5*n uL forward primer
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! Row heading 1
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#: 2.5*n uL reverse primer
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| Cell 2 || Cell 3
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#: 29.5*n uL PCR water
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|-
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# Add 48 uL master mix to each of seven tubes.
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! Row heading A
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# Label tubes blank, 0, 1, 2, 4, 8, 10
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|Cell B
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# Add DNA and MgCl2 to tubes according to chart:
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|Cell C
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<center>{{:Team:Arizona State/Templates/PCR table}}</center>
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|}
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# Add 0.5 uL Phusion polymerase to each tube.
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# Place tubes in thermocycler.
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* Add 0.5 uL Phusion polymerase to each tube.
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# Adjust settings as needed.
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* Place tubes in thermocycler.
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* Adjust settings as needed.
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}}
}}

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.