Keck Subbasement:Schedule

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Week 1

The goal of week 1 is to do a basic experiment so that you will become familiar with the lab and standard molecular biology procedures. We will be exchanging yellow fluorescent protein (YFP) for cyan fluorescent protein (CFP) on a plasmid and then quantifying the results with microscopy.

Monday

  • Pick colonies and grow o/n
  • Vector NTI discussion of two plasmids
  • Design colony PCR/sequencing primers

Tuesday

  • Miniprep plasmid pNNS2-ySMc
  • Amplify CFP
  • Check amplification on a gel
  • Miniprep plasmid pZE12-YFP
  • Digest vector (pZE12-YFP) and insert (YFP PCR product)

Wednesday

  • Gel extraction (vector)
  • PCR purification (insert)
  • Ligation
  • Transformation into competent cells (PM)

Thursday

  • Colony PCR to check for correct insertion
  • PCR purify and prepare product for sequencing
  • Online databases??

Friday

  • Send sequencing
  • Make agarose pads
  • Microscopy
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