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