Keck Subbasement:Schedule: Difference between revisions
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==Week 1== | ==Week 1== | ||
The goal of week 1 is to do a basic experiment so that you will become | 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=== | ===Monday=== | ||
*Pick colonies and grow o/n | *Pick colonies and grow o/n | ||
*Vector NTI discussion of two plasmids | *Vector NTI discussion of two plasmids | ||
*Design colony PCR/sequencing primers | *Design colony PCR/sequencing primers | ||
===Tuesday=== | ===Tuesday=== | ||
*Miniprep plasmid pNNS2-ySMc | *Miniprep plasmid pNNS2-ySMc | ||
*Amplify | *Amplify CFP | ||
*Check amplification on a gel | *Check amplification on a gel | ||
*Miniprep plasmid pZE12-YFP | *Miniprep plasmid pZE12-YFP | ||
*Digest vector (pZE12-YFP) and insert (YFP PCR product) | *Digest vector (pZE12-YFP) and insert (YFP PCR product) | ||
===Wednesday=== | ===Wednesday=== | ||
*Gel extraction (vector) | *Gel extraction (vector) | ||
Latest revision as of 11:11, 24 June 2008
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
