Notebook

March 2016 to March 2017:

1. We work with BioBricks of previous years (kit iGEM 2015. 2016).
2. We re-suspend genetic material and start to do all the standard methods, like
3. The results are not what expected. We cannot see the material in the gel, doing impossible to prove our biological circuit.

April 2017-July 2017

1.We received the distribution kit 2017.
2.We prove again with all the procedure, but we cannot have the results that expected.

July 11th-13th

1.Review and later shipment of parts to IDT to synthesize (glocks Gene Fragments)
2. The parts were:

Saturday 19th, August

1.Re-suspension of the gBlocks (20ng/ul)
2.Digestion of parts. (1 hour for 37°C)

Digestion Protocol: (One enzyme)

1-H2O Mq                      16,3 ul
2-Buffer MC                   2    ul 
3-BSA                             0,2 ul 
4-DNA                             1   ul 
5-Enzyme                      0,5 ul
                   Total: 20ul

Digestion Protocol: (Two enzyme)

1-H2O Mq            15,8 ul
2-Buffer MC          2    ul  
3-BSA                     0,2 ul 
4-DNA                     1   ul 
5-Enzyme 1           0,5 ul 
6-Enzyme 2           0,5 ul 
                   Total: 20ul

Digestion Protocol: (Backbone)

1-H2O Mq              15,8 ul 
2-Buffer H              2    ul  
3-BSA                     0,2 ul
4-DNA                     1   ul
5-Enzyme PstI       0,5 ul  
6-Enzyme EcoRI    0,5 ul_ 
                   Total: 20ul

Wednesday 23rd

1-Ligation. (for 3 hours at 37°C)

Ligation Protocol: (gblock + backbone)

1-H2O Mq            2 ul 
2-Gblock              4ul 
3-Backbone         2ul 
4-Buffer ligase     1ul 
5-T4 Ligase           1ul​
                   Total: 10ul

Saturday 26th

1-Ligation of complete sensor + Lux 2-Digestion