September/5 September 2009

From 2009.igem.org

1.Colony check

sample No.  (no. of colonies)   
 No.5               10  
 No.9               0
 No.18              0
 No.20              0
 No.22              1
 No.23              2
 No.24              0
 No.25              4
 No.27              0

ligation check and transformation again

2. Restriction digest for ligation

K204028

Vector		Insert
2-18H	5	2-16N	10
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2 Buffer	2	No.2	2
dH₂O	11	dH₂O	6
total	20uL	total	20uL

K204029

Vector		Insert
2-18F	5	2-16L	10
EcoRI	1	EcoRI	1
XbaI	1	SpeI	1
No.2	2	No.2	2
dH₂O	11	dH₂O	6
total	20uL	total	20uL

K204030

Vector		Insert
2-18F	5	1-19B	5
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2 Buffer	2	No.2	2
dH₂O	11	dH₂O	11
total	20uL	total	20uL

K204027

Vector		Insert
1-6I	8	No.19	8
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2 Buffer	2	No.2	2
dH₂O	7	dH₂O	7
total	20uL	total	20uL

↓
37°C, 3hr
Agarose Gel Electro-phoresis

No.27-I,V , No.28-I,V , No.29-I,V , No.30-I,V ,ladder

gel cut
purification

3.lagation check

No.9-A,B,C,D , vector , 18-A,B,C , 26 , vector , 5 , vector , ladder
No.18 , 5 , 26 : ligaton OK

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