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Contents 1 Lab 20/08/09 1.1 Metal Sensing Team 1.1.1 Introduction 1.1.2 Practical Outline 1.1.3 Procedure 1.1.3.1 Mini-Preps for the E.coli containing BBa_J33206

Lab 20/08/09

Metal Sensing Team

Introduction

In yesterday's lab session, the metal sensor team inoculated 3 tubes of 3ml LB with the three transformant E.coli cultures grown on the LB + ampicillin plates (i.e. the E.coli bacteria which had taken up the BBa_J33206 DNA). In addition, 0.5ml of each of these three solutions was used to innoculate 3 flasks containing 50ml LB each. The initial inoculations of 3ml LB solutions is for today's mini prep; the subsequent inoculations of 50ml LB solutions is for today's midi prep.

Today, the team hope to firstly carry out a mini prep for the three transformant cultures. Secondly, it is intended that restriction enzyme digests are carried out on a sample of all three culture plasmids along with DNA gel electrophoresis to determine whether the transformed DNA is in fact the BBa_J33206 BioBrick. Thirdly, if time permits, the team hope to carry out a midi prep of the plasmids for later cloning. If all of these steps do not get completed today, they can resumed tomorrow.

Practical Outline

The Metal Sensing team hope to carry out the following tasks today:

1. Conduct a mini-prep of the plasmids contained within the three transformant E.coli cultures (i.e. plasmid pSB1A3 containing BBa_J33206).
2. Carry out restriction digests on these plasmids using EcoRI and PstI.
3. Run both the undigested and digested plasmids on agarose gel in DNA gel electrophoresis.
4. Conduct midi-preps of the plasmids if gel results are positive.

Procedure

Despite having plans to carry out all four procedures listed above, only the following occurred today:

• The mini-preps were conducted
• Restriction enzyme digests were carried out.
• 0.8% Agarose gel made up.

Mini-Preps for the E.coli containing BBa_J33206

To carry out this procedure Dr. Aldridge's Mini-Prep protocol was used.