Difference between revisions of "Team:WHU-China/Experiments"

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<tr><td align="center" valign="middle">PEG-P</td><td align="center" valign="middle">400g/L PEG-6000 in 1x SMM</td><td align="center" valign="middle">Autoclave at 121℃ for 11 min</td></tr>
 
<tr><td align="center" valign="middle">PEG-P</td><td align="center" valign="middle">400g/L PEG-6000 in 1x SMM</td><td align="center" valign="middle">Autoclave at 121℃ for 11 min</td></tr>
 
<tr><td align="center" valign="middle">SMMP</td><td align="center" valign="middle">2x AB3 : 2×SMM= 1:1 Separately</td><td align="center" valign="middle">sterilized</td></tr>
 
<tr><td align="center" valign="middle">SMMP</td><td align="center" valign="middle">2x AB3 : 2×SMM= 1:1 Separately</td><td align="center" valign="middle">sterilized</td></tr>
<tr><td align="center" valign="middle">Solution A</td><td align="center" valign="middle"></td><td align="center" valign="middle"></td></tr>
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<tr><td align="center" valign="middle">Solution A</td><td align="center" valign="middle">206g/L Sucrose, 13g/L MOPS, 1.2g/L NaOH in dH2O.Adjust to pH 7.3 with NaOH </td><td align="center" valign="middle">Filtration</td></tr>
<tr><td align="center" valign="middle"></td><td align="center" valign="middle">206g/L Sucrose, 13g/L MOPS, 1.2g/L NaOH in dH2O.</td><td align="center" valign="middle">Adjust to pH 7.3 with NaOH Filtration</td></tr>
+
 
<tr><td align="center" valign="middle">Solution B</td><td align="center" valign="middle">14.04g/L Agar, 0.70g/L Casamino acids, 35.09g/L Yeast Extract in dH2O</td><td align="center" valign="middle">Autoclave at 121℃ for 15 min</td></tr>
 
<tr><td align="center" valign="middle">Solution B</td><td align="center" valign="middle">14.04g/L Agar, 0.70g/L Casamino acids, 35.09g/L Yeast Extract in dH2O</td><td align="center" valign="middle">Autoclave at 121℃ for 15 min</td></tr>
 
<tr><td align="center" valign="middle">8x CR5 salts</td><td align="center" valign="middle">2g/L K2SO4, 80g/L MgCl2•6H2O, 0.4g/L KH2PO4, 17.6g/L CaCl2 in dH2O</td><td align="center" valign="middle">Autoclave at 121℃ for 15 min</td></tr>
 
<tr><td align="center" valign="middle">8x CR5 salts</td><td align="center" valign="middle">2g/L K2SO4, 80g/L MgCl2•6H2O, 0.4g/L KH2PO4, 17.6g/L CaCl2 in dH2O</td><td align="center" valign="middle">Autoclave at 121℃ for 15 min</td></tr>

Revision as of 19:34, 29 October 2017

Materials

SolutionsIngredientsSterilization
2x AB3(Antibiotic Medium No.3)35g/L AB3(DIFCO) in dH2OAutoclave at 121℃ for 20 min
2×SMM 40mM Maleic acid, 80mM NaOH, 40mM MgCl2•6H2O, 1M Sucrose in dH2O Filtration
PEG-P400g/L PEG-6000 in 1x SMMAutoclave at 121℃ for 11 min
SMMP2x AB3 : 2×SMM= 1:1 Separatelysterilized
Solution A206g/L Sucrose, 13g/L MOPS, 1.2g/L NaOH in dH2O.Adjust to pH 7.3 with NaOH Filtration
Solution B14.04g/L Agar, 0.70g/L Casamino acids, 35.09g/L Yeast Extract in dH2OAutoclave at 121℃ for 15 min
8x CR5 salts2g/L K2SO4, 80g/L MgCl2•6H2O, 0.4g/L KH2PO4, 17.6g/L CaCl2 in dH2OAutoclave at 121℃ for 15 min
12% Proline 120g/LL-proline in dH2OFiltration
20% Glucose200g/L Glucose in dH2OFiltration
CR5-top agar(2.5ml) 1.25ml Solution A, 288μl CR5 salts, 125μl 12% Proline, 125μl 20% Glucose, 713μl Solution B(42℃)Separately sterilized and mix in order
Lysis buffer100mM Na3PO4, 5mg/ml Lysozyme. Adjust to pH 6.3 with H3PO4. 1μl 1 M MgSO4 solution and 2 μl HS-Nuclease* (250 U/μl, cat.# GENUC10700-01, final concentration of 500 U/ml)Filtration

(Other basic molecular biological materials are not listed here)

Attribution of others

1.General Support

    Generally, our project has gained many advice and encouragements from many people, including professors from many different schools of Wuhan University(WHU). College of Life Sciences(CLS-WHU) gave us primary support in every aspect of our project.

2.Project support and advice

    Our project is fully supported Primary PI Prof. Xie, Second PI Prof. Chen, Teacher Long Yan and CLS. During the whole process of establishing and optimizing our project, we got a plenty of precious advices from Prof. Guo Lin, Prof. Yin Lei, Prof. Tang Xiaofeng, Prof. Xie Zhixiong, Prof. Chen Yu, Pro. Cao Zhijian Prof. Liu Qingzhen Associate Prof. Huang Yuping and Associate Prof. Peng Fang of CLS-WHU, researcher Qiu Dongru, and researcher He Feng of Institute of Hydrobiology, Chinese Academy of Sciences(IHCAS), Prof. Liu Tiangang of School of Pharmaceutical Sciences, Wuhan University(SPS-WHU), Prof. Mao Xuhui of School of Resource and Environmental Sciences, Wuhan University(SRES-WHU), a Coach of NCTU, graduates of Prof. Xie’s research group, Prof. Chen Xiangdong’s research group and A.Prof. Huang’s research group, undergraduate Bie Yuanyuan, Wu Lin and Gao Guoming of CLS-WHU.

3.Fundraising help and advice

    Our project is fully funded by CLS-WHU and we mainly received advice about fundraising from Prof. Xie, Prof. Chen and Mr. Long.

4.Lab support

    Our experiments were mainly carried out in Scientific Training Room and Room 6132-6135 of CLS. Experimental materials and instruments are mostly provided by CLS, Prof. Chen’s research group and China Center for Type Culture Collection(CCTCC). Wuhan AuGCT DNA-SYN Biotechnology Co., Ltd synthesized primers and sequenced DNA samples for us. BGI helped us in synthesizing rdhANP.

5.Difficult technique support

    We learned electroporation from Zhang Xiaoming, a graduate of Prof. Chen Xiangdong’s research group and conducted soil sample testing by HPLC in testing center of WHU.

6.Project advisor support

    Two advisors gave us advices in different aspects of our project. Advisor Bie Yuanyuan helped us in actual experiments by suggesting and Advisor Wu Lin offered guidance on personnel management and Jamboree.

7.Wiki support

    Our wiki is entirely completed by ourselves. Nearly every team member is involved in its construction and its final completion is achieved by members of Group of Wiki (See Team Members). Additionally, 7 undergraduates of CLS, including Xin Yuan, Tang Daijiao, Gao Huiqing, Zheng Zhizhen, Wang Xiwei, Liu Yuyan and Zhang Wenhui, helped us in translating Chinese protocols in English.

8.Presentation coaching

    Our presentation is all done by ourselves, not receiving any coaching from any other people.

9.Human Practices support

    Our Human Practices are mostly supported by CLS-WHU.

    There are many other people involved in our project, such as those strangers we‘ve interviewed and surveyed and iGEMers from teams Though its hard to express our sincere thanks to them here in a specific way, we are still really grateful for their warm-hearted help.