Team:Kyoto/Notebook

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Revision as of 14:17, 14 August 2012

August 2

Mutation of FT

Inverse PCR
10xBufer	2mM dNTPs	primer fwd	primer rev	template	polymerase	MilliQ	Total
5	5	1.5	1.5	0.5	1	35.5	50

94°C

Dpn1 Digestion
PCR product	Dpn1
50	2

37℃,1h incubate

Self-ligation
product	MilliQ	Ligase	T4 Kinase	Total
2	7	5	1	15

August 10

Golden Gate Assembly method This method helps us to constract some genes quickly and we can design the order of constractions. We are trying to construct a gene cycle composed of 6 genes( pSB1A3, lacI, GFP, RFP, GFP, DT). After this experiment is confirmed to work, we will so some experiment to check how the numbers of promorters influences the strength of transcription.

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@@ Line 1: / Line 1: @@
+==August 2==
+'''Mutation of FT'''
+{|class="wikitable"
+|+Inverse PCR
+!10xBufer!!2mM dNTPs!!primer fwd!!primer rev!!template!!polymerase!!MilliQ!!Total
+|-
+|5||5||1.5||1.5||0.5||1||35.5||50
+|}
+°C
+{|class="wikitable"
+|+Dpn1 Digestion
+!PCR product!!Dpn1
+|-
+|50||2
+|}   37℃,1h incubate
+{|class="wikitable"
+|+Self-ligation
+!product!!MilliQ!!Ligase!!T4 Kinase!!Total
+|-
+|2||7||5||1||15
+|}
 ==August 10==
 Golden Gate Assembly method
 This method helps us to constract some genes quickly and we can design the order of constractions.
 We are trying to construct a gene cycle composed of 6 genes( pSB1A3, lacI, GFP, RFP, GFP, DT). After this experiment is confirmed to work, we will so some experiment to check how the numbers of promorters influences the strength of transcription.
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