August/9 August 2009

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1.colony check

 ( sample No.)  (no. of colonies)
       9                50<
       24                0 
       29                0
       30                0
       31                8
       32                2
        
        9                1

2.digestion and ligation

digestion with restriction enzyme

K204024

Vector		Insert
terminater(1-23L)	6	No. 14-A	12
EcoRI	1	EcoRI	1
XbaI	1	SpeI	1
No.2	2	No.2	2
dH₂O	10	dH₂O	4
total	20uL	total	20uL

K204029

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

K204030

Vector		Insert
2-18F	10	1-19B	6
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2	2	No.2	2
dH₂O	6	dH₂O	10
total	20uL	total	20uL

↓
37degree 3hr
↓
Agarose Gel Electro-phoresis

No. 24 , 29 , 30 , 10-D , 10-E , ladder

↓
16 degree O/N

3.Min prep
Result of Nanodrop concentration check

 (sample No.)      (concentration)
     1-1D                 27.5
     10-D                 85.1
     10-E                 117.2

4.transformation

ligation sample No. 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40A ,40B
sample No. 24 , 29 , 30

5.digestion and ligation

digestion with restriction enzyme

K204041

Vector		Insert
1-1D	12	1-12M	8
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2	2	No.2	2
dH₂O	4	dH₂O	8
total	20uL	total	20uL

K204042

Vector		Insert
1-1D	12	1-8I	8
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2	2	No.2	2
dH₂O	4	dH₂O	8
total	20uL	total	20uL

↓
37degree 3hr
↓
Agarose Gel Electro-phoresis

No. 41 , 42 , ladder

↓
16 degree O/N