08/09/2014

Heeeyy!! Today is digestion day! We’ll digest Bioremediation Biobrick with EcoRI and XbaI to linearize and the Essential Biobrick with EcoRI and SpeI to release the insert need.We used the following system:

The digestion lasted 2 horas at 37°C. The amount used in the gel was 10ul for each sample and 40 ul was kept frozen at -20°C to be purified later.

Eletrophoretical profile of the digested samples:

1. Bioremediation Biobrick not digested

2. Bioremediation Biobrick digested with EcoRI and XbaI

3. Essential Biobrick not digested

4. EssentialBiobrick digested withEcoRI and SpeI

♥ OMG!!It looks so pretty! ♥

Analysing the profile we concluded that sample number 2 is linearized (and pretty!) and that sample number 4 released a fragment of 1.234 base pairs (even prettier!).

We ran the rest of the micro liters from the digestion (40ul) in another gel just to get the DNA purified.

We isolated the DNA we wanted through the agarose gel 0,8% and we purified it. After we analyzed the purification,analyzedthe eletrophoretical profile of the samles of the Bioremediation Biobrick digested with EcoRI and XbaI and the Essential Biobrick digested with EcoRI and SpeI. We applied in the 0,8% agarose gel 10ul of the sample.

Eletrophoretical profile of the samples,digested and purified:

1. Bioremediation Biobrick linearized with EcoRI and XbaI

2. EssentialBiobrick digested with EcoRI and SpeI.

We could analyze through the eletrophoretical profile that the samples have the size we were looking for!!! Unfortunately, we lost a bit of the DNA in the purification used in the gel, mainly in the Bioremediation Biobrick sample.

Now all we have to do is ligate!!!!! Tomorrow is debate day to develop the strategy for the ligations of our biobricks!!!