Haynes Lab:Notebook/SynBERC Scholars 2013/2013/12/17: Difference between revisions

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Part 1: PCR
'''Part 1: PCR'''
* Use the Open PCR machine to amplify segment from "CMV" plasmid DNA
* Use the Open PCR machine to amplify segment from "CMV" plasmid DNA
* Program name = "First Trial 121713"
*


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# Template DNA only
# Template DNA only
# Primers only
# Primers only
'''Part 2: Fluorimeter'''
* Fluorimeter had to be set up on one 96-hole PCR tube rack + two lids
* Zazu's camera phone was propped up in the black camera cradle with a blue lid from a 50-ml conical tube
# Diluted 100 uL PCR reactions by adding 500 uL molecular bio grade water (600 uL final volume, 6x dilution)
# On hydrophobic slide, applied 80 μL of diluted PCR sample, added 80 uL ready-to-use SYBR green dilution (from DNA day - provided by Dr. Garcia/ Rene Davis)
# Did this three times for sample #1 (all components) and sample #2 (forward primer + template DNA)
# Will continue with other samples after winter break




Revision as of 15:13, 17 December 2013

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12/17/13

  • First trail run of OpenPCR & fluorimeter


Part 1: PCR

  • Use the Open PCR machine to amplify segment from "CMV" plasmid DNA
  • Program name = "First Trial 121713"
Reagent 1 2 3 4 5
2x PCR mix 50 50 50 50 50
P0001 1 1 0 0 1
P0002 1 0 1 0 1
DNA template 0.5 0.5 0.5 0.5 0
N.F. H2O 47.5 48.5 48.5 49.5 48
Final vol 100 100 100 100 100


  1. All components
  2. Forward primer + template DNA
  3. Reverse primer + template DNA
  4. Template DNA only
  5. Primers only


Part 2: Fluorimeter

  • Fluorimeter had to be set up on one 96-hole PCR tube rack + two lids
  • Zazu's camera phone was propped up in the black camera cradle with a blue lid from a 50-ml conical tube
  1. Diluted 100 uL PCR reactions by adding 500 uL molecular bio grade water (600 uL final volume, 6x dilution)
  2. On hydrophobic slide, applied 80 μL of diluted PCR sample, added 80 uL ready-to-use SYBR green dilution (from DNA day - provided by Dr. Garcia/ Rene Davis)
  3. Did this three times for sample #1 (all components) and sample #2 (forward primer + template DNA)
  4. Will continue with other samples after winter break



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