TUDelft/9 September 2008

=September 9th=

3A Assembly
A further construction of our parts has been started, we cut all the thermosensitive parts already received and started a 3A assembly in a pSB1AK3 vector of K115012 and K115021 - K115028 (if possible). Ligation was performed in a bit better conditions as last time (stoichiometrically). However the ligation was not o/n but in 2 hours at 16ºC.

Thermosensitive DNA
Another vector with our thermosensitive DNA arrived today, which has been transformed into TOP10 directly again.

PCR
We did another PCR of the E. coli genes with Taq polymerase. Picture of the gel bands which were cut out can be found below.



DNA was isolated from the gel bands and yielded the following concentrations: atoB - 19ng/ul; idi - 18ng/ul; ispA - 18ng/ul. All 260/280 readings were above 1.7, which is associated with pure and proper double stranded DNA.