August/6 October 2009

From 2009.igem.org

1. coloony check

sample no.    no. o f colony
  59                        0
  61                        0
  59                        1
  61                        4
  64                       ++
  65                        6
  X5                        10
  X6                        +
  X7                        10
  X8                        10
 1-24E                    50

2. min prep

sample      conc.
  58            174 ng/uL
  60            220
  62            267
  63            237 
  64            151
  65            75
  67            252
  68            233
  X3           236
  X4           193

3.transformation

sample no. 
11 , 20 , 21 , 2-6O , 1-15J , 1-15L

4.digation
K204052

Vector		Insert
1-6I	10	33	6
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2	2	No.2	2
dH₂O	6	dH₂O	6
total	20uL	total	20uL

K204067

Vector		Insert
1-23L	6	2-16L	10
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2	2	No.2	2
dH₂O	10	dH₂O	6
total	20uL	total	20uL

K204067

Vector		Insert
1-23L	6	2-16L	10
EcoRI	1	EcoRI	1
XbaI	1	SpeI	1
No.2	2	No.2	2
dH₂O	10	dH₂O	6
total	20uL	total	20uL

K204068

Vector		Insert
1-23L	10	1-19B	5
EcoRI	1	EcoRI	1
XbaI	1	SpeI	1
No.2	2	No.2	2
dH₂O	6	dH₂O	11
total	20uL	total	20uL

↓
37°C, 2hr
gel cut & purification
ligation O/N

5.prepared LA pate

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