Formal Lab Session - 23rd September 2009

Team Newcastle 2009 iGEM 23-09-09 IMG 1358.JPG

Overview

Stochastic Switch Team -

Sporulation Tuning/Chassis Team -

Stochastic Switch Team

Team Newcastle 2009 iGEM 23-09-09 IMG 1346.JPG

Today we discovered that the transfrormations seemed sucessful, and the control plates had no growth at all which is what was hoped for. Hopefuly this means that we have a transformed ara-sspb-pSB1AT3 ligation product.

We set up miniprep cultures for 12 colonies using LB + amp + tet
We also set up 12 more miniprep cultures for the Sac transformed cells in hope that the precious minipreps had been carried out incorrectly.

Sporulation Tuning/Chassis Team

Team Newcastle 2009 iGEM 23-09-09 IMG 1340.JPG

Introduction

Today's main work is to ligate kinA with pGFP-rrnB vector and transform ligated vector into E.coli.

Experiment procedure

ligate kinA and pGFP-rrnB

ligate reaction

 dd H2O                                 14.5ul
 Fast ligation buffer                      4ul
 pGFP-rrnB backbone(180ng/ul)            0.5ul
 kinA fragment(6.1ng/ul)                  15ul
 T4 ligase                                 1ul
 ---------------------------------------------
                                          35ul

Votex for few seconds.
22C 10min and 1 hour in RT.

Digest pMK-RQ:kinA and pGFP-rrnB

pMK-RQ digest reaction.

 dd H2O                     7ul
 10X fast digest buffer     7ul
 Fast EcoRI                 3ul
 Fast NheI                  3ul
 pMK-RQ vector             50ul
 ------------------------------
                           70ul

pGFP-rrnB digest reaction.

 dd H2O                     7ul
 10X fast digest buffer     7ul
 Fast EcoRI                 3ul
 Fast NheI                  3ul
 pGFP-rrnB vector          50ul
 ------------------------------
                           70ul