August/20 August 2009

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(New page: 1.<B>digestion and ligation</b><br> digestion with restriction enzyme<br> '''K204007''' , (K204003) <table border="1" Frame="box"> <tr><th colspan="2">Vector</th><th colspan="2">Insert</...)
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Revision as of 05:33, 22 August 2009

1.digestion and ligation

digestion with restriction enzyme

K204007 , (K204003)

Vector		Insert
RBS(1-2M)	10	cinR(1-14F)	12
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2 Buffer	2	No.2	2
dH2O	6	dH2O	4
total	20uL	total	20uL

K204008

Vector		Insert
RBS(1-2M)	10	LuxI(2-8O)	12
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2 Buffer	2	No.2	2
dH2O	6	dH2O	4
total	20uL	total	20uL

↓
37°C, 2hr

↓
gel electrophoresis
gel cut
↓
purification by [QIAquick Nucleotide Removal Kit]
↓
ligation

ligation
DNA        44
10* buffer 5
ligation   1
total      50uL
↓
16°C overnight

2. rapid check

③(RBS+cinR),K204003

③-C,D : OK

④(RBS+LasR),K204004

④-A,B,C : OK

⑥(RBS+cinI),K204006

⑥-A,B,C : OK