September/10 September 2009

From 2009.igem.org

1.colony check

  ( sample No.)  (no. of colonies)
      24                7
      24                0
      29                1
      30                0
      30                0
      33               100<
      34                50<
      35                2
      36                0
      37                0 
      38                0
      39                1
      40               10

2.transformation

ligation sample No. 41 , 42 , 14 , 22
3-1M

37degree 10hr

3.Min prep
Result of Nanodrop concentration check

(sample No.) (concentration)
     9A             16.3
      B             13.4
      C             21.8
      D             15.3
     
     31A            17.4
       B            14.3
       C            15.3
       
     32A            14.5
       B            11.9

4.digestion and ligation

digestion with restriction enzyme

K204043 : 1-16I

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

K204044 :1-1K

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

↓
gel purification
↓
ligation 37degree O/N

5.ligation check

UP   : 10-D , 10-E , 9-A , 9-B , 9-C , 9-D , ladder
DOWN : 31-A , 31-B , 31-C , 32-A , 32-B , ladder

No.9 OK!

6.transformation

ligation sample No. 24 , 29 , 30 , 36 , 37 , 38 , 43 , 44

7.colony check

 ( sample No.)  (no. of colonies)
      3-1M             0 
       14             +++
       22             +++
       41             +++
       42             ++

back to NOTES