August/25 August 2009

From 2009.igem.org

1.trandformation

ligation sample No. 12 ,13 , 14 , 15 
↓
65°C , 10min
↓

competent cell 100uL
     sample     10uL 

competent cell 100uL
     sample      2uL

2.Min prep
Result of Nanodrop concentration check

(sample No.) (concentration)
    1-6I         13.5
                    19.1
                    15.4
                    13.4
                    18.9
     
    1-4E         15.0
                    15.3
                    14.9
                 
    2-16N        22.7
                    17.7
                    23.1
       
    2-16H        13.8
                    22.7
    
    2-16L        22.5
                    31.5

3.ligation check

sample No. 9 , 10

4.digestion
digestion with restriction enzyme

K204019

Vector		Insert
RBS(1-2M)	8	2-13N	6
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2 Buffer	2	No.2	2
dH₂O	8	dH₂O	10
total	20uL	total	20uL

K204020

Vector		Insert
RBS(1-2M)	8	YFP(1-8I)	6
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2 Buffer	2	No.2	2
dH₂O	8	dH₂O	10
total	20uL	total	20uL

K204021

Vector		Insert
RBS(1-2M)	8	CFP(1-8K)	6
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2 Buffer	2	No.2	2
dH₂O	8	dH₂O	10
total	20uL	total	20uL

↓
37°C, over night

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