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Team:Tsinghua
From 2008.igem.org
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Reagent Concentration/Activity 50ul 100ul | Reagent Concentration/Activity 50ul 100ul | ||
| - | |||
GF taq buffer 10x 5 10 | GF taq buffer 10x 5 10 | ||
GF taq 0.5 0.5 | GF taq 0.5 0.5 | ||
| Line 33: | Line 32: | ||
Primer 1 10uM 1 2 | Primer 1 10uM 1 2 | ||
Primer 2 10um 1 2 | Primer 2 10um 1 2 | ||
| - | Template DNA | + | Template DNA changeable 0.5 1 |
MgCl2 0.2M 0.5 1 | MgCl2 0.2M 0.5 1 | ||
ddH2O --- 40.5 81 | ddH2O --- 40.5 81 | ||
| Line 41: | Line 40: | ||
Progress Program I Program 2 | Progress Program I Program 2 | ||
| - | |||
Predenaturing 95℃ 2-5 min 95℃ 2-5 min | Predenaturing 95℃ 2-5 min 95℃ 2-5 min | ||
Denaturing 95℃ 10-20sec 95℃ 10-20sec | Denaturing 95℃ 10-20sec 95℃ 10-20sec | ||
| Line 52: | Line 50: | ||
Reagent Concentration/Activity Volume(50ul system) | Reagent Concentration/Activity Volume(50ul system) | ||
| - | |||
Restriction cut buffer 10x 5ul | Restriction cut buffer 10x 5ul | ||
Enzyme 1 -- 2.5ul | Enzyme 1 -- 2.5ul | ||
| Line 62: | Line 59: | ||
3 Ligation: | 3 Ligation: | ||
Reagent Volume(10ul system) | Reagent Volume(10ul system) | ||
| - | |||
Solution I 5ul | Solution I 5ul | ||
DNA fragment 3.5ul | DNA fragment 3.5ul | ||
Revision as of 10:15, 28 July 2008
Although this page is still in heavy construction, every guest is welcomed to left your trace on the "Doodle Board".
Our Team Project Parts Experiments Communication Links Doodle Board
Our Team
Luying JiaYuanfan Yang Gechong Ruan Shan Lin Xin Rong Yicheng Long
Ziying Liu Yuemeng Wang
Yilong Zou
Zi Wang
He Yang
Yongqiang Gou Chao Wang
Junjie Luo Cong Liu
Project
This year we are trying to work at the motility of bacteria. E.coli senses some chemicals and moves toward or stays still, this behavior is called chemotaxis. Any modification of the pathway from the sensor to the motile apparatus will cause into different chemotactic effcts.
Parts
Communication
Experiment and Protocol
1 PCR
1.1 Gold Faster Taq system:
Reagent Concentration/Activity 50ul 100ul
GF taq buffer 10x 5 10
GF taq 0.5 0.5
dNTPmix 10mM each 1 2
Primer 1 10uM 1 2
Primer 2 10um 1 2
Template DNA changeable 0.5 1
MgCl2 0.2M 0.5 1
ddH2O --- 40.5 81
1.2 The program under Gold faster taq system
Progress Program I Program 2 Predenaturing 95℃ 2-5 min 95℃ 2-5 min Denaturing 95℃ 10-20sec 95℃ 10-20sec Annealing (Tm-5) ℃ 2-5 sec 68℃ 10-15sec/1kb Extension 72℃ 10-15sec/1kb
25-30cycle 25-30cycle
Last extension 1-2min or skipped 1-2min or skipped
2 Restriction cut:
Reagent Concentration/Activity Volume(50ul system)
Restriction cut buffer 10x 5ul
Enzyme 1 -- 2.5ul
Enzyme 2 -- 2.5ul
Incubate at 37℃, 1.5 hrs or longer (Enzymes from Takara Co., Ltd)
3 Ligation:
Reagent Volume(10ul system)
Solution I 5ul
DNA fragment 3.5ul
Vector 1.5ul
Incubate at 16-18℃,1hr or longer (Ligation kit from Takara)
Links
Tsinghua University: http://www.tsinghua.edu.cn/qhdwzy/index.jsp
Parts: http://partsregistry.org
iGEM Headquarter: hq@igem.org
Beijing Normal University:https://2008.igem.org/Team:Beijing_Normal
ETH_Zurich:https://2008.igem.org/Team:ETH_Zurich
Nature:http://www.nature.com
Science:http://www.sciencemag.org
System and Synthetic Biology:http://www.springer.com/biomed/journal/11693
Doodle Board
Oh, My E.coli, where are you! Yilong 08.7.11
