Lab Work - 08/09/09

Metal Sensing Team

Introduction

Yesterday's lab session saw the Metal Sensing team innoculate 6 tubes of 5ml LB (plus ampicillin) with six cultures of DH5-alpha transformed with BBa_J33206 in plasmid pSB1A2 in the morning. By the time afternoon had arrived, the bacteria had grown to sufficient numbers; three of these cultures were then used to further inoculate 3 flasks of 50ml LB+amp. The three flasks of 50ml LB+amp + transformant E. coli cells were then placed in the orbital incubator and will take part in today's midi-prep. Meanwhile the six tubes were used in a mini-prep attempt but alas, the procedure went wrong and had to be abandoned.

Today's lab session will focus on creating midi-prepped plasmids from the inoculated 50ml LB+amp flasks as well as carry out restriction enzyme digest analysis on a sample. The samples will then be run on agarose gel by DNA gel electrophoresis.

Procedure

Midi-prep of transformant E. coli cells

To carry out the midi-prep we used Sigma-Aldrich's GenElute midi-prep kit and with it, the protocol. There were no changes to the protocol and a summary steps can be seen below. The only thing to note is that we did not midi-prep all of the samples placed in the flask; cultures 1 and 2 (both deriving from the 200ul LB+amp plates) were both spun down with supernatants removed (culture 2 from the 500ul plate was discarded) with culture 1 being placed in the freezer for storage and culture 2 used for the midi-prep process.

Restriction Enzyme digests

DNA gel electrophoresis analysis

Results

20 – 21 June 2009 - Europe workshop (London)
23 – 24 June 2009 - UK iGEM meetup (Edinburgh)
23 October Practice Presentation (Newcastle)
23 October T-shirts are ready
27 October Practice Presentation (Sunderland)
27 October Poster is ready
30 October – 2 November 2009 - Jamboree (Boston)

Newcastle iGEM Twitter
[http://www.facebook.com/home.php#/group.php?gid=131709337641 Newcastle on Facebook]
[http://www.youtube.com/user/newcastle2009igem Newcastle Youtube Channel]

Team:Newcastle/Labwork/8 September 2009

From 2009.igem.org