From 2007.igem.org

07/23/07

• We observed the uncorrected grew of I13507 on the plate and none of R0051.
• We got along with:

- another transformation of R0051 (2 ul), I13507 and C0051;

- the ligation of J04500+J04631 (I763004) to test the ligation protocoll with 2 experiences: vector and insert 2 ul, vector and insert 2-4 ul. We leave it at 25°C for an hour, then at 65°C for 10 minutes.

• Then we transformed 4 ul of ligate.

07/24/07

• We observe colony for C0051 on the plate. Then we preinoculate in 5 ml all the day and at evening we inoculate in 50 ml and we leave at 37°C O/N.
• We don’t observe any colony for R0051 and I13507 and for the 2 ligations.

GFP induction test in J04431 transformed cells: we preinoculate in 5 ml. At OD: 0.4/0.6 + IPTG, we put a drop on the micro slide. Yet after 30 minutes the cells are green.

• We harvest them and we put them in 5 ml without IPTG to see the extinction time of the protein.
• We read every 20 minutes.

• We get along with another GFP induction: yet after 10 minutes the cells are green.
• We transform bacteria wiht 4 ul of I13507 and R0051 and with 10 ul of J04500 + J04631 (I763004) ligations.
• We plate after we have harvested.

07/25/07

• We get along with Miniprep for C0051:

-Quantification: conc. 65 ng/ul;

-Cleanness n.:1.8.

• We observe colony for I13507 and ligation. We preinoculate in 5 ml and at evening in 50 ml.
• We don’t find any coloby for R0051.
• We transform with all the R0051 we have and alternatively with R1051.

07/26/07

07/27/07

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