User:Tara K. Luckau/Notebook/Team ConGen/2010/10/29

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Thoughts and Musings

DNA Concentration

  • Tina is prepping mass DNA extraction for optimization (using extra samples with lots of chunky)
  • Want to use these samples to figure out relative DNA concentration to use for all future PCR optimizations
  • Take Tina's extractions and spot checks
  • Run PCR on DNA serial dilution; gel for which concentrations produced product
  • note smallest concentration that worked
  • shove it through NanoDrop
  • Compare that sample's spot check and NanoDrop - that's the brightness of spot we'll want for all future PCRs

Questions

  • since I don't really have a verified working primer set, how can I expect the PCR to work?
  • will necessarily need to run DNA concentration gradient, temp gradient and buffer gradient in same reaction



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