Haynes Lab:Notebook/SynBERC Scholars 2013/2014/02/13: Difference between revisions

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==Entry title==
==Procedure Digital PCR==
* Insert content here...
 
'''Part 1'''
 
*TNF A forward and reverse primers were diluted from 100uM to 10uM. Initial stock DNA (Concentration = 2.3mg/mL) was diluted several times to obtain 4 different templates, listed below:
*U20S 1:100 = 6.24ng/mL
*D1 = 0.624ng/mL
*D2 = 0.0624ng/mL = 6.24*10^(-2) ng/mL
*D3 = 6.24pg/mL
 
 
 
*For each PCR reaction, the following volumes will be added:
* 1uL of forward TNF A primer
* 1uL of reverse TNF A primer
* 1uL of diluted DNA (either U20S 1:100, D1, D2, or D3 for each sample)
* 50uL of PCR master mix
* 47 uL of dH20
 
*8 samples of each template will be put through the PCR  sequence (NOTE: The program to be run is named "First Trial 121713")
*2 samples of plain water will also be tested as a control.  




Revision as of 15:56, 13 February 2014

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Procedure Digital PCR

Part 1

  • TNF A forward and reverse primers were diluted from 100uM to 10uM. Initial stock DNA (Concentration = 2.3mg/mL) was diluted several times to obtain 4 different templates, listed below:
  • U20S 1:100 = 6.24ng/mL
  • D1 = 0.624ng/mL
  • D2 = 0.0624ng/mL = 6.24*10^(-2) ng/mL
  • D3 = 6.24pg/mL


  • For each PCR reaction, the following volumes will be added:
  • 1uL of forward TNF A primer
  • 1uL of reverse TNF A primer
  • 1uL of diluted DNA (either U20S 1:100, D1, D2, or D3 for each sample)
  • 50uL of PCR master mix
  • 47 uL of dH20
  • 8 samples of each template will be put through the PCR sequence (NOTE: The program to be run is named "First Trial 121713")
  • 2 samples of plain water will also be tested as a control.



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