1. We work with BioBricks of previous years (kit iGEM 2015. 2016).
1.We received the distribution kit 2017.
1.Review and later shipment of parts to IDT to synthesize (glocks Gene Fragments)
Notebook
March 2016 to March 2017:
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
2.We prove again with all the procedure, but we cannot have the results that expected.
July 11th-13th
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