September/21 September 2009

From 2009.igem.org

1.colony check

 sample no.  no. of colony
   27            ++
   28            ++
   30            ++
   35            + 
   45            +
   46            1
   
   48            ++
   50           +++
   51            +
  1-1F          +++
   
   46            1
  2-9A           +
  2-3I           ++
  3-7G           ++
  3-7C           ++
  1-8K          +++

2.transformation

sample no
36 , 37 , 38 , 39 , 40

3.min prep

sample no.     conc.
  27           285 ng/uL
  28           181
  30           194
  35           173
  45           290
  48           35
  50           153
  51           146
 1-1F          132

4.digation

X1-A

Vector		Insert
51	4	1-12M	8
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2	2	No.2	2
dH₂O	12	dH₂O	8
total	20uL	total	20uL

X2-A

Vector		Insert
50	4	1-12M	8
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2	2	No.2	2
dH₂O	12	dH₂O	8
total	20uL	total	20uL

X3-A

Vector		Insert
31	16	1-12M	4
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2	2	No.2	2
dH₂O	0	dH₂O	12
total	20uL	total	20uL

X4-A

Vector		Insert
32	16	1-12M	4
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2	2	No.2	2
dH₂O	0	dH₂O	12
total	20uL	total	20uL

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

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