Keck Subbasement:Schedule

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==Week 1==
==Week 1==
 +
The goal of week 1 is to do a basic experiment so that you will become framiliar with the lab and standard molecular biology procedures. We will be exchanging yellow fluorescent protein (YFP) for cyan fluorescent protein on a plasmid and then quantifying the results with microscopy.
 +
===Monday===
===Monday===
-
*Pick colony and grow o/n
+
*Pick colonies and grow o/n
*Measure OD growth curves
*Measure OD growth curves
-
*Design colony PCR primers
+
*Vector NTI discussion of two plasmids
 +
*Design colony PCR/sequencing primers
===Tuesday===
===Tuesday===
-
*Miniprep fluorescent protein (FP)
+
*Miniprep plasmid pNNS2-ySMc
-
*Amplify FP
+
*Amplify YFP
-
*Digest vector and insert
+
*Check amplification on a gel
-
*Miniprep large quantity of promoter plasmid
+
*Miniprep plasmid pZE12-YFP
 +
*Digest vector (pZE12-YFP) and insert (YFP PCR product)
===Wednesday===
===Wednesday===
*Gel extraction (vector)
*Gel extraction (vector)
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*Transformation into competent cells (PM)
*Transformation into competent cells (PM)
===Thursday===
===Thursday===
-
*Colony PCR to check correct insert
+
*Colony PCR to check for correct insertion
-
*PCR purify and design primers
+
*PCR purify and prepare product for sequencing
*Online databases??  
*Online databases??  
===Friday===
===Friday===

Revision as of 12:47, 18 June 2008

Contents

Week 1

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

Monday

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

Tuesday

  • Miniprep plasmid pNNS2-ySMc
  • Amplify YFP
  • 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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