Team:FJNU-China/Notebook
An unforgettable journey
Week1 27/10/2017
First meeting, Team FJNU-China was established.
Week2 03/2-10/2
Learn the molecular biology experimental techniques under the guidance of Dr.Yang.
Week3-Week7 Apr
Sent d-ldh, Tyrb and rocG to the company for synthesis. Obtained d-ldh, Tyrb and rocG fragment by PCR.[Result1] [Result2][Result3]
![Obtained d-ldh, Tyrb and rocG fragment by PCR.[Result1]](https://static.igem.org/mediawiki/2018/1/17/T--FJNU-China--PCR_Tyrb.png){kind=link}
![[Result2]](https://static.igem.org/mediawiki/2018/b/ba/T--FJNU-China--pcr_D-ldh.jpg){kind=link}
![[Result3]](https://static.igem.org/mediawiki/2018/8/81/T--FJNU-China--Fig.8_3.1_rocG_pcr_KZ_.jpg){kind=link}
Week8-Week12 May
1.Extract genome of E. coli Bw25113.2.Construct d-ldh, Tyrb and rocG by Gibson assembly. 3.PCR verification. [Result]
![[Result]](https://static.igem.org/mediawiki/2018/3/3d/T--FJNU-China--Fig.8_3.2_rocG_pcr_YZ_.jpg){kind=link}
Week13-Week17 June
1.Extracted plasmid of E. coli BM4R 2.Cultured Clostridium acetobutylicum 3.Induction culture of engineering bacteria 4.SDS-PAGE verifies protein expression [Result]
![[Result]](https://static.igem.org/mediawiki/2018/7/72/T--FJNU-China--SDS-PAGE_Dldh-Tryb-rocG.jpg){kind=link}
Week18 1/7-7/7
1.Synthetic primer for our experiments. 2.Cultured staphylococcus epidermidis. 3.Did a PCR pre-experiment to amplify atoD, atoA 4.Verified that if E.coli can grow without salt. 5.Held a summer camp with Quanzhou 7th High school and Fuzhou 8th High school.
![[Result]](https://static.igem.org/mediawiki/2018/d/db/T--FJNU-China--16spcr.png){kind=link}
![[Result]](https://static.igem.org/mediawiki/2018/5/5d/T--FJNU-China--atoD-atoA.png){kind=link}
![[Result]](https://static.igem.org/mediawiki/2018/4/42/T--FJNU-China--mazF-backbone.png){kind=link}
![[Result]](https://static.igem.org/mediawiki/2018/5/5e/T--FJNU-China--PCR-lacI.png){kind=link}
Week22 22/7-28/7
1.Attended the iGEM Eurasian Meetup 2018 2.Verified the function of MazF with different concentrations of arabinose concentration.[Result] 3.Did the T4 ligation for lacI and the circuit we synthesized. 4.Added different concentrations of PLA to samples and Measured OD value. 5.Tried to obtain eGFP fragment by PCR. 6.Uploaded the data of InterLab.
![[Result]](https://static.igem.org/mediawiki/2018/5/57/T--FJNU-China--arabinose-induced-MazF.png){kind=link}
Week23 29/7-04/8
1.Making a standard curve of PLA. 2.Failed to constructed the circuit of hns-syn-eGFP, we decided to replace the eGFP with the GFP. 3.Change the backbone of the plasmid hns-syn from PUC57 into pRB1S. 4.Add restriction sites for atoD, atoA by PCR. 5.Did the lethal experiment of PLA against Staphylococcus epidermidis to provide data for our model.
Week24 5/8-11/8
1.Failed to enzyme cutting pRB1s, PLA and mazF with Kpn1 and Dra3. 2.Failed to construct pRB1s-atoD-atoA by Gibson assembly. 3.Obtained GFP fragment by PCR. 4.Tried to construct pRB1s-hns-syn-GFP. 5.Obtained ARO10, par and TyrB fragment by PCR. [Result] 6. Construct ARO10, par, TyrB and pSB1C3 by Gibson assembly.
![[Result]](https://2018.igem.org/File:T--FJNU-China--ARO10-par-TyrB.jpg){kind=link}
Week25 12/8-18/8
1.Constructed pSB1C3-atoD and pSB1C3-atoA by T4 connection. 2.Communicated with XMU-China. 3.The lethal experiment of mazf against Staphylococcus epidermidis. 4.Site-directed mutation.
Week26 19/8-25/8
1.The lethal experiment of mazf against Staphylococcus epidermidis. 2.T4 ligation for GFP and PLA. 3.Measured the production of 2-PE and PLA
Week27 26/8-01/9
Attended 5th ccic in Shanghai,China.
Week28 2/9-8/9
1.Got the fragment of adhE2. 2.Cultured our engineered bacteria in 0M, 0.05M, and 0.3M at 25℃ and 37℃.
Week29 9/9-15/9
1.Construct pRB1s-atoD-atoA-adhE2 and composite part. [Result] 2.T4 ligation for salt-temperature circuit and pSB1C3. 3.Line cultured the hns-syn on the different salt concentration of plates.
![[Result]](https://static.igem.org/mediawiki/2018/8/89/T--FJNU-China--atoa-atod-adhe2.jpg){kind=link}
Week30 16/9-22/9
1.Observed the GFP level under fluorescence microscope.[Result] 2.verify whether circuit is successfully constructed. 3.Construct improve part.
![[Result]](https://static.igem.org/mediawiki/2018/5/54/T--FJNU-China--fluorescence-microscope.png){kind=link}
Week31 23/9-29/9
Constructed araC+pBAD+tetR+GFP+PLtetO1+RFP part. [Result]
![[Result]](https://static.igem.org/mediawiki/2018/d/db/T--FJNU-China--tetrteto.png){kind=link}
Week29 30/9-07/10
1.Submitted our parts 2.Made wiki 3.Verified circuit function