Haynes Lab:Notebook/CRISPR Editing/2014/08/05: Difference between revisions

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==08/05/2014==
==08/05/14==


* SURVEYOR assay - for mCherry CRISPR
* SURVEYOR assay - for mCherry CRISPR
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* Borun et al used at LEAST 50 ng genomic DNA in their reactions (final amount per PCR ambiguous)
* Borun et al used at LEAST 50 ng genomic DNA in their reactions (final amount per PCR ambiguous)
** We will test 20 ng, 50 ng, and 100 ng
** We will test 20 ng, 50 ng, and 100 ng




Revision as of 07:57, 5 August 2014

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08/05/14

  • SURVEYOR assay - for mCherry CRISPR
  • qPCR melt curve optimization - mCherry CRISPR 1

SURVEYOR assay

PCR primers [Rene, please add this info]

  • Forward = ???, Reverse = ???
  • Amplicon length = ???

PCR templates

  1. CRISPR "1"
  2. CRISPR "2"
  3. CRISPR "4"


qPCR melt curve optimization - mCherry CRISPR 1
Assistance from Kumaran Lleng

Optimization parameters

  • Borun et al reported cleaner peak profiles after a 30-cycle amplification (vs. 40 cycles)
    • We will use 30 cycles instead of 45
  • Borun et al used at LEAST 50 ng genomic DNA in their reactions (final amount per PCR ambiguous)
    • We will test 20 ng, 50 ng, and 100 ng





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