Team:ZJU-China/October.html

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<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd"> Notebook - September

Lab Notes

July August
 September
 October

Protocol

Protocol

Brainstorm

Brainstorm

Lab Notes - October

Biobrick Group

Week13

Day Note
Sept.26th Monday
•Miniprep: Rush-3,-4,-12
•Run the gel of digestion test results:
RFP+YFP+1K3-7,-16		 •PCR amplify: Rush-3,-4,-12
•Sample and test the intensity of fluorescence once or twice an hour during the growth of E.coli
Sept.27th Tuesday
•Ligate and transform:
RFP+YFP+CFP+1C3 RFP+YFP+1C3		 •PCR amplify:20H
•Cut: Rush Bone,22B
Sept.28th Wednesday
•Cultivate: DH5αwith Device CFP,RFP,YFP and without any device
•Ligate and purify:Rush+22B
•Cut and purify:
the product of the ligation		 •Run the gel: Digested Rush and 22B,Lig1
•Self-ligate:Digested Lig
•Colony PCR: RFP+YFP+CFP+1C3
•Cut and purify: RFP+YFP+1K3,12I
Sept.29th Thursday
•Ligate and transform: RFP+YFP+1K3&1C3,
RFP+YFP&1C3
•Colony PCR:RFP+YFP+CFP+1C3
•Preserve the colony on the plate
•Plate the ligation results: RFP+1C3,
RFP+YFP+CFP+1C3
Colony PCR:RFP+YFP+1C3
RFP+YFP+CFP+1C3
•Cultivate the above
Sept.30th Friday
•Colony PCR: Rush-Lig
Oct.1st Saturday
•Colony PCR: Failed
•MIniprep: 10I		 •Cut: 10I+22B,20J,1T3
•Ligate:10I+22B+1T3		 •Cultivate:2I,2M
Oct.2nd Sunday
•Transform the ligation results:
22B+10I(1T3),20J+10I(1T3)
•Miniprep and Preserve:2M,2I		 •Cut:fdhf,Vgb,2M,2I
•Purify:RFP+YFP+CFP+1C3		 •Ligate and transform:
RFP+YFP+CFP&1A3k
fdhf+2M&1K3,Vgb+2I&1K3

Week14

Day Note
Oct.3rd Monday
•Colony PCR
Oct.4th Tuesday
•Cut:RBS+20J
•Cultivate:fdhf+2M-1,-2,-3,-4
Vgb+2I-1,-222B+10I-1,-2
•Ligate and purify:RBS+20J& 1C3 		•Cut:RBS+20J
•Ligate:2M+20J&10I to 1K3
•Transform:2M+20J+10I,Vgb+1C3
•Cultivate:22B+10I-1,-2		 •Miniprep and cut: fdhf+2M,Vgb+2I
•Cut:fdhf+2M,20H
Oct.5th Wednesday
•Cultivate:Vgb+2I 1-6
•Ligate:fdhf+20H& 1K3 		•Transform:fdhf+20H
•Cut:2M+20J
Oct.6th Thurday
Finishing the wiki
Oct.7th Friday
making poster
Oct.8th Saturday
•Miniprep and test:
Vgb,fdhf+20H+1K3 1-4		 •Colony PCR
Oct.9th Sunday
•Digestion test and run the gel to test the results:
20J+10I -1,-2,-4
22B+10I -1,-2,-3,-4 Vgb+1C3
Fdhf+2M+20H -1,-2,-3,-4 Vgb+2I -1,-2,-5		 •Colony PCR:fdhf+2M+20H 5-14
•Cultivate:fdhf+2M+20H 5-14
•Cut:fdhf+2M,20H

 

Week15

Day Note
Oct.10th Monday
•Ligate:Fdhf+2M&20H to 1C3
•Cultivate:16L+Device C		 •Cut:Vgb+2I,22B+10I
•Ligate:Vgb+2I&22B+10I to1C3
Oct.11th Tuesday
•Cultivate:20H,22B+10I-1,20J+10I-1,-4 •Miniprep and cut:
fdhf+2M+20H+1K3 5-14
Oct.12th Wednesday
•Miniprep: fdhf+20H-10,20H,22B+10I, 20J+10I-1,-4 fdhf+2I+20H, •Cut and run the gel: Vgb+2I-5,
22B+10I,20J+10I,1C3 backbone
Oct.13th Thursday
•Cut and run the gel to test the ligate results
Oct.14th Friday
•Regional Jamboree @ HKUST
Oct.15th Saturday
•Regional Jamboree @ HKUST
Oct.16th Sunday
•Regional Jamboree @ HKUST We are the Champion!!!

 

Week16

Day Note
Oct.17th Monday
•Rest
Oct.18th Tuesday
•Rest
Oct.19th Wednesday
•Transform: Kit Plate 1 18E&18I, Kit Plate 4 7K& 7M
Oct.20th Thursday
•Transform: Kit Plate 4 7K
•Colony PCR and run the gel:23L 1-6
Oct.21st Friday
•Transform:7K
•Cultivate:7M,18I,18E,23L pSB1A3,1K3,1C3 backbone		 •Cut and test the resistance: 22M+10I
•Colony PCR:7M 1-7,18E 1-6,18I 1-6
Oct. 22nd Saturday
•Cultivate:pSB1C3-5,23L-1,-2,7M-1
•Cultivate:7K,10I,22M,18E,18I
•Colony PCR:18E 1-6,18I 1-6
•Test the reagent for PCR several times		 •Miniprep and digestion test: 7M,23L,1K3,1A3,1C3
•Digestion and run the gel: 7M,22M+10I-2,1K3
Oct.23rd Sunday
•Cut:7M,22M+10I
•Digestion test:18I,18E		 •Ligate and transform: 7M+22M+10I& 1K3

Week17

Day Note
Oct.24th Monday
•Transform: Kit Plate 4 8G,8I Kit Plate 1 1F
•Culture yeast & B. subtilis		 •Colony PCR and run the gel: 7M+22M+10I+1K3 1-25
•Cultivate:8G,1F,8I
Oct.25th Tuesday
•Miniprep:7M+22M+10I+1K3
•Colony PCR:8I,8G,1F
•Cut:7M+22M+10I+1K3,1C3 backbone		 •Cultivate:7K
•Ligate:7M+22M+10I& 1C3
•Genome DNA extraction from yeast & B. subtilis
•PCR condition pre-test for alsS & Adh2
Oct.26th Wednesday
•Cultivate:8G,8I,1F,7M,18E,18I
•Transform:7M+22M+10I+1C3 7M+22M+10I+1K3-2
•Reserve:7M+22M+10I+1K3,8G,8I, 1F,7M,18E,18I		 •Miniprep:8G
•Cultivate:7M+22M+10I+1C3，2J
•Testify PCR results on gel
•Clean-up alsS PCR product
•Re-pre-test PCR condition for Adh2
Oct.27th Thursday
•PCR producing Adh2
•alsS ES double-digestion
•purification both
Oct.28th Friday
Oct. 29th Saturday
Oct.30th Sunday


Biofilm Group

3rd Oct

18 Oct

Biofilm formation of AI2+rainbofilm device without induction of AI2 for 48 hours.

19 Oct

Co-transformation of AHL and rainbofilm device.

20 Oct

Observation of rainbofilm. Maximum thickness is 14 micrometer.

22 Oct

Biofilm formation with or without AI2 (by arabinose induction of part BBa_K091191) for 48, 36, 24 hours.

24 Oct

Laser confocal observation of biofilm. Used too much water to wash away salt crystals and although thick biofilm is observed before washing but very thin biofilm is observed under laser confocal.

25 Oct

Biofilm formation with rainbofilm DH5alpha without quorum sensing for 48 hours. Colony PCR of 20 JM109 AI2-rainbofilm co-transformaion. Non has rainbofilm plasmid successfully transformed.

26 Oct

18 AI2-rainbofilm JM109 and 20 AHL-rainbofilm DH5alpha colony PCR. Number 5,6,14 of AI2 and number 5,8,11 of AHL have both plasmids.

27 Oct

Biofilm formation with AI2, AHL induced in rainbofilm DH5alpha AHL, DH5alpha AI2 and JM109 AI2.

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