Team:CIEI-China/Notebook/Lab Book

Jan. 2017

We came up with the idea about using salt tolerant yeast to dispose kitchen waste which have high osmotic pressure. Then we started to search the papers related to our research.

Mar. 2017

We found three genes that are possible for our goal: SpTPS1, ScTPS1, and gltB.

Apr.2017

And then we began to prepare the materials we need in the following experiments.

May. 2017

We finished the design of nine biobricks with different functions.

Jun. 2017

Designing and editing of the experiment plan.

Transformation of PUC57 plasmid into DH5α.

Culture the E.coli.

Preparation of plasmid DNA by Alkaline Lysis with SDS;

Digestion (XbaⅠ, SpeⅠ).

Recycle the electrophoresis product.

Linearization of the standard carrier.

Week 1-2 in Jul. 2017

Ligate the genes into the standard carrier.

Transform the standard carrier into DH5α E.coli;

Culture the E.coli and prepare the plasmid.

Week 3 in Jul. 2017

Design the primers of the three genes: SpTPS1, ScTPS1, and gltB.

Week 4 in Jul. 2017 to week 1 in Aug. 2017

PCR PrimeStar DNA polymerase.

Recycle the plasmids from PCR products.

Digest the PCR products.

Electrophoresis and recycling.

Week 2 –week 3 in Aug.2017

Ligation;

We successfully ligated the three target genes in to the vector pPIC9K to get the devices.

Transform the vector into DH5α E.coli.

Culture the E.coli.

Extract the devices.

Week 4 in Aug. 2017 to week 1 in Sept. 2017

Electrophoresis of six completed devices: intracellular and extracellular expression of the three target genes. Recycle the devices from the gels.

Transform the devices into GS115 competent yeast cells.

Week 2-4 in Sep. 2017

Sequencing the genes we transformed into the yeast.

Induce experiments by using methanol.

SDS-Page.

Week 1-3 in Oct. 2017

Test the survive rate of the yeast in different salinity.

Test the decomposition rate of the waste.