• Home
• Team
• Project
• Parts
• Modelling
• Sponsors
• Lab Book

Project

• Overview
• Choices Rationale
• Characterisation
• Safety
• Judging Comments

Sub-projects

• Metal Intake/Efflux
• Metal Sensing
• Sporulation Tuning
• Population Dynamics
• Stochastic Switch
• Metal Sequester
• Chassis
• Promoter Library

Wet Lab

• Lab Book
• Protocols

Meetings

• Meetings Calendar

Planning

• Timeline
• Ideas

Links

• Helping other teams
• Ethics
• T-Shirts
• iGEM Meetup
• Our City
• Fun and Games
• Sponsors
• Useful Links
• Contact Us
• Press Coverage

Contents 1 Lab Work 04/09/09 1.1 Promoter Library Sub-Project 1.1.1 Introduction 1.1.2 Preparing samples and DNA Gel Electrophoresis 1.1.3 Results

Lab Work 04/09/09

Promoter Library Sub-Project

Introduction

In yesterday's lab session the products from the PCR reactions on the C0, V1, V2 and V3 samples were purified. The gel electrophoresis carried out on the unpurified samples the previous day showed hazy bands however it was uncertain that these bands were the promoters or just primer-dimers. Today, we aim to carry out DNA gel electrophoresis on these purified samples (in 1.5% agarose gel) to determine whether any PCR amplification reactions worked.

Preparing samples and DNA Gel Electrophoresis

The purified PCR products were treated in a similar way to which DNA samples are treated in the gel electrophoresis protocol except 5ul of each sample was extracted and to each of these 1ul of loading buffer (dye) added. 15ul of HindIII' ladder will be added in the first and last wells of the 1.5% agarose gel. This is how the wells will be organised:

• Lane 1 = blank
• Lane 2 = 15ul of HindIII DNA ladder
• Lane 3 = 6ul of C0 (Sample 1)
• Lane 4 = 6ul of V1 (Sample 2)
• Lane 5 = 6ul of V2 (Sample 3)
• Lane 6 = 6ul of V3 (Sample 4)
• Lane 7 = 15ul of HindIII DNA ladder

The 1.5% agarose was prepared and poured in the same way as the DNA gel electrophoresis protocol suggests. The samples were then added to the wells in the way described above; the voltage was set to 60mv and left for 15-20 minutes. The photograph obtained from the resulting gel can be seen in the 'Results' section.

Results