Team:PKU Beijing/Notebook/AND Gate 1/Input/LuxR

Notebook > AND Gate 1 > Input > HSL Sensor

2009.6.9 (with LinMin)
11:10 The e-coli cultivated yesterday did not grow The material of the former culture medium went bad. Need renewing.

11:29 Practise minipreping the plasmids

12:58 Practise digesting the plasmids

2009.6.13 (with LinMin)
15:00 make the culture medium, repair computer, search Parts:

16:55 dissolve the plasmids, prepare to transform add 15uL ddH2O

21:20 Transform plasmids above

2009.6.14
13:10 repair the computer

14:08 make the LB culture medium

21:00 search the rbs parts:

2009.6.15
21:07 Transform plasmids above (2009/06/14)

21:48 begin to transforming

2009.6.16
11:50 cultivation is finished The results come out well

2009.6.19 (with Wu Shuke)
21:11 Select clones from the plate

2009.6.20
23:00 transform 1-12L

2009.6.21
9:50 Transform the 2008 Parts’ RBS

11:54 finish transformaion

2009.6.22
9:14 Transformation is failed, redoing

22:30 re-transformation

2009.6.23
1:00 start the cultivation

13:00 the second transformation is failed

2009.6.24
14:59 Prepare to make TOP10 Sensitive Cells

16:00 Begin to make LB, etc

17:30 Search for LuxR s

23:38 Cultivate TOP10 Cells

2009.6.25
21:44 Transform the rbs: 1003-12C; 1003-12G; 1004-2A; 1004-2C; 1004-3G

2009.6.26
22:36 Cultivate: 1004-1C; 1004-3C; 1004-4C; 1004-2G

23:07 Prepare to make 1-2I Sensitive Cells

23:15 Cultivate 1-2I Cells

2009.6.27
15:58 Miniprep the plasmids: 1004-1C; 1004-3C; 1004-4C; 1004-2G Failed

16:09 Make the 1-2I Sensitive Cells Failed

2009.6.28
20:18 Miniprep the plasmids: 1004-1C; 1004-2C; 1004-3C; 1004-4C; 1004-2G

21:37 Make the 1-2I Sensitive Cells

2009.6.29
17:40 Check the 1-2I Sensitive Cells Huge Success! The protocol is added to ftp

20:09 Prepare to make DH5A Sensitive Cells

2009.7.3
18:00 Search Parts:

22:44 Cultivate 1003-6C Failed. The plasmid is bad.

2009.7.3-2009.7.10
Searching for papers on T3P and T3Pol, which is basic for the second logic gate. Contact with the writer to get the plasmids. During this time, a review on T7 and T3 systems is summarized, which has been sent to ftp.

2009.7.6
19:51 Miniprep the plasmids:

21:08 Digest the plasmids of 2-4O and 1-18P:

2009.7.7
0:34 The digestion reaction begins

7:46 add 0.4uL cip and 2ul buffer to the vector system

8:07 the result come out as follows:

18:03 Recycle the fragments.

19:40 Ligate the 2-4O insert to 1-18P vector:

19:48 Ligation start

23:44 Begin the transformation of the ligation plasmid

2009.7.8
16:57 Cultivate the 2-4O->1-18P

2009.7.9
9:06 Miniprep the 2-4O->1-18P plasmids

11:52 Digest the plasmids of 2-4O->1-18P and 1-23L:

12:13 Reaction Start

17:32 Stop the digestion

20:08 Prepare to Ligation

20:17 Ligation start