• 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 14/08/09 1.1 Metal Sensor Team 1.1.1 Introduction and Summary 1.1.2 Work we did

Lab 14/08/09

Metal Sensor Team

Introduction and Summary

So far we have shown that our Bacillus subtilis 168 cultures, once the GFP-rrnb DNA has been added, can survive on LB + Chloramphenicol media. This suggests that we have successfully transformed our B. subtilis cells as the wild type B. subtilis cells do not have this resistance. However we need to prove that the resistance inherited by the Bacillus subtilis bacteria is due to the GFP-rrnb and not acquired by other means. Today, we intend to pour some LB + Chloramphenicol + starch plates and then plate our 'transformed' Bacillus cells onto these plates. The reason for adding starch to the plates: the GFP-rrnb plasmid removes the Bacillus subtilis bacteria's ability to break down starch using the amyE enzyme - starch plates will prove the bacteria have been transformed.

Work we did

• Using the 'pouring plates' protocol, we made up 1 litre of LB + agar + starch solution and once it had been autoclaved and properly prepared (with chloramphenicol - this is the resistance displayed in GFP-rrnB), the solution was poured into plates.

• These plates were then left on the bench for 1 hour. Three plates were taken from the stack and placed in the 42C incubator to dry off.