Team:Peking S/lab/notebook/sr

• Rui Sun's Notebook

Summary

In this summer, I mainly focused on the construction of the microbial population density balancer. In addition, I also carried out the assay for orthogonality of positive regulating signaling molecules.

Contents

• July, 2011

• August, 2011

• September, 2011

• October, 2011

July

[TOP]

7.7

ABBREVIATIONS:

Restriction endonuleases:

E=EcoRI X=XbaI S=SpeI P=PstI

17:30 Digestion of B0034(1-2M) with S&P and 4-12I with X&P started.

     Fragment lengths: B0034  around 2000bps
                       4-12I  756 bps
     PCR of terminator-less version of Ptet-rhlI and lacI(I732011)-LVA

     Electroperesis:
     Gel 1 Well 2-4: Marker_4-12I_1-2M
     Gel 2 Well 2-5: Marker_rhlI_rhlI_rhlI
     Gel 3 Well 1-6: Marker_lacI_lacI_ _ _lacI_lacI
     All checked right.

     Gel extraction of the above DNA fragments.

     Ligation of B0034(SP digested)+4-12I(XP digested).

7.8

12:30 Transformation of the ligation product(1-2M+4-12I) finished. 12:30 Gel extraction of PCR product (LacI-LVA) finished.

17:30 PCR for lacI-tag from I732100 started.

19:30 Transformation of 2-10G finished.

19:30 PCR failed. Repeated with the same system &condition and got the right string.

7.9

0 :19 Checked plates.

0 :19 Gel extraction & PCR of lacI-LVA-suffix.

14:00 Digestion of 1-1G and lacI PCR product(second round) with X&P.

14:00 Colony pick of 1-1G plates.

18:30 Electrophoresis for digestion products(1-1G XP fragment;lacI PCR product digested with X&P).

23:45 Ligation of 1-1G and lacI-suffix fragments started.

7.10

00:05 Digestion of Ptet-rhlI PCR products (1 and 2) started. 00:05 Digestions: 2-10G(EX) 1-2M+4-12I(RBS+gfp-LVA)(1,2,3，ES) 1-23L（EX&ES) started.

7.12

7.13

7.14

7.15

7.16

7.17

7.18

7.19

7.20

7.21

7.22

7.23

7.24

7.25

7.26

7.27

7.28

7.29

7.30

7.31

August

[TOP]

8.1

8.2

8.4

8.5

8.6

8.7

8.8

8.9

8.10

8.11

8.12

8.13

8.14

8.15

8.16

8.17

8.18

8.19

8.20

8.21

8.22

8.23

8.25

8.26

8.27

8.30

8.31

September

[TOP]

9.1

9.2

9.3

9.6

9.7

9.8

9.9

9.12

9.13

9.14

9.15

9.16

9.17

9.21

9.22

9.23

9.24

9.25

9.26

9.27

9.28

9.29

9.30

October

[TOP]

10.1

10.3

10.4

10.5

10.7

10.8

10.9

10.10

10.11

10.12

10.13

10.15

10.16-10.21

10.21-10.25