"
Uppsala-Sweden/13 August 2009
From 2009.igem.org
Transformation of ADH2 from z.mobilis, continued
Unfortunatly we only got two (!!) colonies from Adh2 z. These are most likely to be contamination as one was found on 50 µl olate and on on the 550 µl plate, we will still evaluate them with a colony PCR however.
The control pUC 19 yielded 59 colonies at the 20 µl plate giving a transformation efficiency of 70,8*10^6 cfu/µg pUC19. This compared to what is considered good transformation efficiency by [http://openwetware.org/wiki/TOP10_chemically_competent_cells TOP10 chemically competent cells], 500 - 5000 *10^6 cfu/µg pUC19.
Becouse of this we decided to test the competence of our cells against another batch of the same type of cells done here in the lab. This is done today and incubated over nigth so we will have the answer tomorrow.
If the other cells are as crappy we will change to TOP10 cells also available. The most likely couse of failure is that we autoclaved the CCMB80 buffer, against protocol, with coused a reddish brown, likely manganese, precipitation.
When transforming this test we also throw in some Adh2 z to see if we get lucky. Even with very bad cells (and ours are merly not very good it seems) there should be some colonies of Adh2 z.We investigate the couse of this below.
--Anders 15:44, 13 August 2009 (UTC)
