Notebook
Week 1
Wet lab
We choose the plasmid of kit plate to transform and amplify. We design parts by adding promoter (BBa_K206000) and RBS(BBa_B0034) to the upstream region of luciferase gene and adding prefix and suffix and and we committed the GENEWIZ company to synthesize this gene.
We gather and summarize the data of algal bloom case in North China.
Week 2
Wet Lab
We transformed the plasmid pSB1C3(BBa_K592009/ BBa_E1010/BBa_J04450) in DH5α and proceeded amplification, extracted the plasmid and used two kinds of enzyme (EcoR I,Pst I) to digest and verify.
We designed primers and committed the GENEWIZ company to synthesize these primers(nluc、nluc-psb1c3、cluc、cluc-psb1c3、β-tubulin、β-tubulin-psb1c3、α-tubulin、α-tubulin-psb1c3).
We borrowed expression plasmid pET21a (+) from the lab of Yang Dong to amplify and DNA template (nluc、cluc、mcf-7、hepg-2),and we amplified these fragments (nluc、cluc、α-tubulin、β-tubulin) by PCR.
Dry Lab
We guide FAFU about modeling.
Week 3
Wet lab
We designed primers about topo(α-tubulin-TOPO-F、β-tubulin-TOPO-F、α-pET21a(+)、β-pET21a(+)、nluc- pET21a(+)、cluc -pET21a(+))and committed the GENEWIZ company to synthesize these primers.
Testing amplification products by PCR, and linked them to produce plasmids PSB1C3 and plasmids pET21a(+). Meanwhile we did topo clone.
We did PCR amplification by using efficient polymerase, did restriction enzyme (EcoR I/Xho I) digestion and ligation about PCR products(α-TOPO、β-TOPO、α-pET21a(+)、β-pET21a(+)、nluc- pET21a(+)、cluc -pET21a(+)).
Dry Lab
We modified basic logistic growth curve combined with specific growing environment of Algae through reviewing literatures about Algae growthing modeling.