August/7 October 2009

From 2009.igem.org

1. colony check

sample no.     no. of colony
   11                   +++
   20                   +++
   21                   +++
  2-6O                 ++
  1-15J                +
  1-15L                +

2.transformation

sample no.
67 , 68 , 69 , 52 , 11 , 20 , 21 , 2-6O , 1-15J , 1-15L

3.digation

K204070

Vector		Insert
33	2	50	5
EcoRI	1	EcoRI	1
XbaI	1	SpeI	1
No.2	2	No.2	2
dH₂O	12	dH₂O	9
total	20uL	total	20uL

K204071

Vector		Insert
1-8K	2	51	4
EcoRI	1	EcoRI	1
XbaI	1	SpeI	1
No.2	2	No.2	2
dH₂O	12	dH₂O	10
total	20uL	total	20uL

K204072

Vector		Insert
2-8E	2	31	6
EcoRI	1	EcoRI	1
XbaI	1	SpeI	1
No.2	2	No.2	2
dH₂O	12	dH₂O	12
total	20uL	total	20uL

K204073

Vector		Insert
1-8I	6	32	6
EcoRI	1	EcoRI	1
XbaI	1	SpeI	1
No.2	2	No.2	2
dH₂O	8	dH₂O	8
total	20uL	total	20uL

K204074

Vector		Insert
1-1K	10	1-12M	2
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2	2	No.2	2
dH₂O	4	dH₂O	12
total	20uL	total	20uL

K204075

Vector		Insert
1-1K	10	1-8K	2
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2	2	No.2	2
dH₂O	4/td>	dH₂O	12
total	20uL	total	20uL

↓
37 degree . O/N

4. min prep

sample            conc.
 X4                   257 ng/uL
 X4                   266
 X4                   171
 X5                   169
 X6                   293
 X7                   294
 X8                   151
 59                   306
 61                   285
 64                   252
 65                   278

5.digation

X4

Vector		Insert
32	16	1-12M	4
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2	2	No.2	2
dH₂O	2	dH₂O	12
total	20uL	total	20uL

↓
37 degree , 2hr
gel cut

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