IGEM:UNAM-Genomics Mexico/2009/Notebook/H2 Pae/2011/06/22

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Doing Competent cells We centrifugate the cells which we left yesterday on CaCl and we eliminated the supernatant. We added glycerol, and we divided all the cells into 20 ependorf tubes, containing 50 micro litters each. With this the competent cells of E. Coli BL21_DE3 were ready. Transformation With the cells previously created I did the transformation of the plasmid pBBRMCS-5, which was isolated by Pato from an E. coli DH5-alfa. The transformation was done accordanly this protocol, but with just 1 tube of 50 microliters and no control, this because Miguel told me to do it in that way. At nigth I came to see if colonies had grown, and there were around 25 colonies so Pato incubated 4 colonies in 5 ml of liquid medium(Gm) each of them, in order to isolate pBBRMCS-5 tomorrow.