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Contents 1 Lab Session 27/07/2009 1.1 Running the gel for RFP and GFP bioricks 1.2 Transformation of new biobricks 1.3 Preparation of the plates 1.3.1 Tomorrow

Lab Session 27/07/2009

Running the gel for RFP and GFP bioricks

Today we prepared the gel with 0.8% agorose for last week's rescued plasmids which contain RFP and GFP biobricks. The final solution was 500ml using 4 gr of agorose in total.

We mixed 9ul of DNas with 1ul of the buffer and centrifuged shortly. The rest of the DNAs are then put back to the freezer.

Finally we run the gel. We loaded the ladder to the first and fourth wells, GFP to the 2nd and RFP to the 3rd well and set the voltage to 120V which was then reduced to 90V.

Below are the images:

Transformation of new biobricks

We prepared the agar plates and left the tubes including the biobricks for incubation overnight.

We checked the agar plates containing colonies of plasmids with five different biobricks from last week. We will be following Phil’s mini method for Alkaline Lysis for Mini Prep The list of plates and the biobricks are below:

Plate	BioBrick	Number used for the labelling	Name
Plate 1	[http://partsregistry.org/Part:BBa_C0056 BBa_C0056]	1	CI Coding Sequence
Plate 2	[http://partsregistry.org/Part:BBa_B1002 BBa_B1002]	2	Terminator
Plate 3	[http://partsregistry.org/Part:BBa_C0077 BBa_C0077]	3	CinR Coding sequence
Plate 4	[http://partsregistry.org/Part:BBa_C0076 BBa_C0076]	4	CinI Coding Sequence
Plate 5	[http://partsregistry.org/Part:BBa_R0077 BBa_R0077]	5	CinR sensitive promoter

Except from plate 3, we had colonies formed in all plates.

Colonies from the plates were picked and mixed in a tube containing 5ml of LB. We prepared 12 tubes for 4 different biobricks. For each biobrick we labelled the tubes as A, B, and C. Finally we left them at shaking incubator overnight.

Preparation of the plates

We prepared the agar solutions and poured into the plates which are then placed into the fridge. The list of plates and their content are as below.

Content	Number of plates prepared	Labelled as
LB	4	LB 27/07/09
LB + Amp	6	LB+A 27/07/09
LB + Amp + Kan	5	LB+AK 27/07/09

Tomorrow

We have placed the required solutions to the bench for the experiment apart from RNAase. We need to find it first thing in the morning tomorrow then we will follow Phil’s mini method for Alkaline Lysis for Mini Prep. GFP and RFP are placed into the freezer. The DNA ladder is in the fridge. Plates are placed into the fridge just below the top shelf. They were split into two since they did not all fit to the shelf.