Notebook

Here is a summery of our notebook.

Protocols

Contants

• Top
• NanoBio Protocol for gene knockout
• Antibiotics preparation
• Chemical transformation
• DNA Kit Plate
• Gel and PCR Extraction
• Gel Prep
• Glycerol stock
• LB SOB SOC BA
• Ligation
• manual NEB (gibson assembly)
• MixandGo
• PCR
• Phosphorylation and blunt ligation
• Presto Mini Plasmid Kit Protocol
• Restriction enzyme
• Transformation to 3300 LG using electroporation
• AHL inducer experiment protocol

Week	Design	Cloning	Experiment	Finish new BioBrick
Jul 5th to Jul 9th (2015)	Primers : OGEM 1 and 2 to open j09855 plasmid . Gblock :RBS- Aiia with a Gibson overlabs.	we got j09855 part from the distribution kit.	-	-
June 12th to July 16th (2015)	Primers : OGEM 3 and 4 to got the k1767001 plasmid without LuxR , after doing the PCR we will get a linear DNA so we put two enzymes in the overlaps of the primers to close the plasmids.	PCR to open the j09855 plasmid.Gibson to add Aiia .	-	BBa_K1767001
Aug. 2nd to Aug. 6th (2015)	-	We got p0440 part from the distribution kit. We built BBa_K1767002 and BBa_K1767004 Constructs by restriction enzymes. PCR to get the k1767001 plasmid without LuxR .	-	BBa_K1767002 BBa_K1767004
Aug. 9th to Aug. 13th (2015)	Primers : OGEM 5 and 6 to check if we succeed in the PCR ( Those primers for colony PCR)	We got I13521 part from the distribution kit. We built BBa_K1767003 and BBa_K1767005 Constructs by restriction enzymes. Colony PCR to check if we have LuxR in the plasmid.	-	BBa_K1767003 BBa_K1767005
Aug. 16th to Aug. 20th (2015)	-	-	AHL experiment #1	-
Aug. 23rd to Aug. 27th (2015)	Primers : OGEM 7 and 8 to check if we have yfp in the plasmid ( for colony PCR ).	We got I13003 part from the distribution kit.	AHL experiment #2	-
Aug. 30th to Sept. 9th (2015)	Primers: OGEM 9 and 10 to replaced the RFP with RFP ( with LVA )	We built BBa_K1767009 and BBa_K1767010 Constructs by restriction enzymes. Colony PCR to check if we have YFP in the plasmid.	-	BBa_K1767009 BBa_K1767010
Sept. 6th to Sept. 11th (2015)	-	-	AHL experiment # 3	-

preparation

• 1. LB medium
• 2. Miniprep

July 12th

July 13th

July 15th

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July 29th

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september 1th

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september 7th