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Contents 1 Introductory Lab Sessions Week 3 1.1 The third week initial planning, 14th July - 17th July 1.1.1 Lab session: 15th July 1.1.1.1 Experiment Recap 1.1.1.2 Introduction 1.1.1.3 Procedure

Introductory Lab Sessions Week 3

(14th July - 17th July 2009)

The third week initial planning, 14th July - 17th July

• Tuesday

1. DNA electrophoresis of restriction enzyme digested plasmids
2. Innoculate LB with two selected cultures (decision based on DNA gel electrophoresis results)

• Wednesday
  • Innoculate a flask of LB with the two selected cultures

• Thurday

• Friday

Lab session: 15th July

Experiment Recap

In the previous experiment, we had carried out DNA gel electrophoresis on six plasmids from our transformed E.coli cells - the six plasmids were each divided into two sets with the first set of six being treated with EcoRI alone and the second set being treated with EcoRI and PstI:
First set - treated with EcoRI

• Culture 2 (GFP) - Lane 3
• Culture 3 (RFP) - Lane 4
• Culture 4 (RFP) - Lane 5
• Culture 5 (RFP) - Lane 6
• Culture 6 (RFP) - Lane 7
• Culture 7 (RFP) - Lane 8

Second set - treated with EcoRI and PstI

• Culture 2 (GFP) - Lane 9
• Culture 3 (RFP) - Lane 10
• Culture 4 (RFP) - Lane 11
• Culture 5 (RFP) - Lane 12
• Culture 6 (RFP) - Lane 13
• Culture 7 (RFP) - Lane 14

After analysis of the gel photographs and comparison with the expected band sizes for the plasmids, plasmid backbones and BioBricks, we decided that cultures 2(GFP) and 3(RFP) were to be used for a larger scale plasmid prep.

To 2 tubes of sterile LB nutrient broth, a sample of cultures 2 and 3 were introduced. They were then placed into the shaking incubator overnight.

Introduction

Today's procedure is straightforward. Using aseptic technique, we are to innoculate 2 flasks containing LB nutrient broth with the 2 cultures grown overnight. Once carried out, these flasks will be incubated overnight and used in tomorrow's larger scale plasmid prep.

Procedure